JP4129722B2 - Petri dish for selective collection of sperm - Google Patents

Petri dish for selective collection of sperm Download PDF

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JP4129722B2
JP4129722B2 JP2002104286A JP2002104286A JP4129722B2 JP 4129722 B2 JP4129722 B2 JP 4129722B2 JP 2002104286 A JP2002104286 A JP 2002104286A JP 2002104286 A JP2002104286 A JP 2002104286A JP 4129722 B2 JP4129722 B2 JP 4129722B2
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petri dish
sperm
recess
selective collection
concave portion
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JP2003289850A (en
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智 兼子
信雄 高木
徳寛 比恵島
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Nipro Corp
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/10—Petri dish
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    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization

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Description

【０００１】
【発明が属する技術分野】
本発明は、精子懸濁液から運動精子を、顕微鏡下にて選択的に採取するために用いられる精子選択採取用シャーレに関する。
【０００２】
【従来の技術】
生殖補助技術（ＡＲＴ）の中でも、重度の精液所見不良例などを対象とする顕微授精（ＩＣＳＩ）は、精子が雌性生殖路を遡上し、卵と受精、融合するまでの全ての過程を体外で行う方法である。この体外受精によれば、射精精液中に精子が極端に少ない症例や、精液中の精子が自力で卵子に侵入する能力（受精能）に欠けた症例であっても、理論的には１個の精子と１個の卵が存在すれば受精が可能となる。ここで用いる精子は、射精精子、精巣上体精子または精巣精子のいずれであってもよい。
【０００３】
従来、ＡＲＴにおいては、精液より取り出された形態的に正常な運動精子を受精に用いることが重要とされている。該運動精子は、密度勾配体を用いる密度勾配遠心分離法により精液を濃縮し精漿を除去した後、精子を含有する密度勾配体（以下、精子懸濁液と呼ぶ）を1〜5mlの培養液中に静置し、自身の運動性により培養液中を上昇（swim up）した精子を回収する方法（swim up法）によって得られる。しかし、ＩＣＳＩの対象となる精液所見不良例は、精液中の精子数が少ない場合や、精子の運動率が低い場合が多く、このようなswim up法では十分な運動精子を回収することができない。
【０００４】
そこで、ＩＣＳＩにおける運動精子の回収には、1〜10mlの精液を密度勾配遠心分離法により10〜50μlに濃縮した精子懸濁液と、50〜100μlの培養液が予め導入されているシャーレとを用い、該培養液中に精子懸濁液を注入した後静置して、該懸濁液中でswim upした運動精子を回収する方法が用いられる。この方法においては、使用する精子懸濁液および培養液が少量であるため、運動精子の回収は顕微鏡下で行うことができる。
しかし、前記ＩＣＳＩにおける運動精子回収方法において用いられるシャーレは、底面が平滑であるため、シャーレ底面に導入した培養液に精子懸濁液を注入すると、自然拡散により該懸濁液が培養液全体に広がるため、運動精子と非運動精子を分離する操作は非常に困難である。
【０００５】
【発明が解決しようとする課題】
上記事情に鑑み、本発明は自然拡散による影響を抑え、運動精子のみを効率よく選択採取することが可能なシャーレを提供することを目的とする。
【０００６】
【課題を解決するための手段】
本発明者らは、上記課題を解決するために種々鋭意検討した結果、従来のシャーレの底面に、ＩＣＳＩに用いるために濃縮された精子懸濁液を収容しうる内容積を有する凹部を形成したことにより、該懸濁液の自然拡散による影響を受けることなく運動精子のみの採取を可能とするシャーレを提供できることを見出した。
【０００７】
すなわち、本発明は、底面と側壁とを有し、該底面に0.01〜0.2mlの容積を有する凹部が設けられてなる精子選択採取用シャーレである。
また、本発明は、底面と側壁とを有し、該底面が段階的に深くなる形状を有し、その最深部が0.01〜0.2mlである容積を有する凹部である精子選択採取用シャーレである。
【０００８】
【作用】
本発明のシャーレは、底面に設けられた凹部に精子懸濁液を注入するため、該懸濁液が培養液中に自然拡散することを抑制し、かつ、自身の運動性により凹部から進出した運動精子のみを選択採取することが容易に行える。また、凹部から進出した運動精子はシャーレの凹部以外の底面上に存在するため、顕微鏡下での識別が容易で、効率よく運動精子を採取することができる。
【０００９】
【発明の実施の形態】
以下に、本発明のシャーレを添付図面に示す好適な実施例に基づいて詳細に説明するが、本発明はこれらの説明に限定されるものではない。
図１は本発明のシャーレの一実施例を示す側面図および断面図であり、図２は本発明のシャーレの他の実施例を示す側面図および断面図である。
【００１０】
本発明の一実施形態を図１を用いて説明する。図１に示すシャーレ１は、底面２と側壁３とからなり、好ましくは円形の底面２を有し、底面２の直径に対してはるかに小さい高さを有する側壁３とからなる有底筒体である。該シャーレ１は、予め精子の生存性を維持するための培養液を収容し、そこに濃縮された精子含有液（後述する精子懸濁液）を注入して、ＩＣＳＩに用いられる運動精子の選択採取を行うものである。したがって、該シャーレ１は前記培養液および精子懸濁液の収納を可能とする内容積を有するものであり、その好ましい寸法は、底面２の直径が30〜120mmであり、側壁３の高さが3〜10mmである。該側壁３の高さが低いほど、内容積が小さくなるが、精子懸濁液の注入および運動精子の採取の動作が行いやすくなる。
前記シャーレ１は、電子線滅菌が可能で、ポリスチレンやポリエチレン、ポリアミドなどの透明性の高い材料で形成されたものであることが好ましい。
【００１１】
前記シャーレ１の底面２には、凹部４が設けられてなる。該凹部４は、ＩＣＳＩ用の運動精子の選択採取に際し、密度勾配体を用いる密度勾配遠心分離法などにより濃縮された、精子を含有する密度勾配体（精子懸濁液と呼ぶ）を収容しうる内容積を有するものである。具体的には、該凹部４の内容積は0.01〜0.2mlであり、好ましくは40〜80μlである。該凹部４の形状は特に限定されないが、好ましくは直径2〜10mm、深さ2〜10mmの有底筒体である。
該凹部４は、シャーレ１の底面２に厚みをもたせて該底面２に埋没するように設けられてもよいし、図１に示すように底面２から下方へ突出するように設けられてもよい。後者の場合、シャーレ１の側壁３は、凹部４の底面よりも下方へ突出する脚部５を形成するものであれば、シャーレ１の戴置が安定して行えてより好ましい。
前記凹部４は、シャーレ１の底面２のどの位置に設けられてもよいが、シャーレ１の成形上および精子濃縮液の注入操作上、底面２のほぼ中央部に設けられることが好ましい。
【００１２】
本発明の他の実施形態を図２を用いて説明する。本発明のシャーレは、底面が段階的に深くなる形状を有するものであってもよく、好ましくは図２に示すような２段階で深くなる底面を有するシャーレ１１である。該シャーレ１１は、底面１２と側壁１３とからなり、好ましくは円形の底面１２を有し、底面１２の直径に対してはるかに小さい高さを有する側壁１３とからなる有底筒体である。該シャーレ１の寸法や材料は図１に示すシャーレ１と同じものが適用される。
該シャーレ１１の底面２には、第一凹部１４１が設けられ、該第一凹部１４１の底面１４２には第二凹部１４３が設けられてなる。該第二凹部１４３は図１に示すシャーレ１における凹部４と同じ役割を果たすものであり、好ましい寸法および形状も同様に規定される。
前記シャーレ１１の底面１２に設けられる第一凹部１４１は、より運動性の高い精子を選択するためのものである。該第一凹部１４１の容積はシャーレ１１の容積よりも小さく、具体的には、底面１４２の直径が20〜80mmであり、深さが2〜5mmである。該第一凹部１４１は有底筒状、すなわち側壁１４４が底面１４２に対して垂直である必要はなく、むしろ側壁１４４が底面１４２に対して角度を有するすり鉢状であることが、精子の選択採取操作上好ましい。
【００１３】
また、前記第一凹部１４１および第二凹部１４３は、シャーレ１１の底面１２に厚みを持たせて該底面１２に埋没するように設けられてもよいし、図２に示すように底面１２から下方へ突出するように設けられてもよい。後者の場合、シャーレ１１の側壁１３は、底面１２よりも下方へ突出する脚部１５を形成するものであればより好ましい。
前記第一凹部１４１および第二凹部１４３は、シャーレ１１の底面１２のどの一に設けられてもよいが、シャーレ１と同様、底面１２のほぼ中央部に設けられることが好ましく、底面１２，第一凹部１４１および第二凹部１４３のそれぞれの底面が全て同心円であることがより好ましい。
【００１４】
次に、本発明のシャーレの使用方法を、図１に示すシャーレ１１を用いて説明する。
まず、シャーレ１の凹部４周辺にＨＴＦ（Human Tubal Fluid Medium）やHuecm（ニプロ社製、登録商標）などの培養液を約50〜100μl導入し、さらに該培養液の蒸発を防止するために、該培養液表面をミネラルオイルなどで被覆する。次いで、密度勾配遠心分離法などの公知の方法により濃縮された精子懸濁液10〜50μmを、注射針、ガラスキャピラールなど先の細い管状の器具を用いて該凹部４内に注入し、静置する。
凹部４内に注入された精子懸濁液は、従来のシャーレを用いた場合と比較して、培養液との接触面積がはるかに小さいため、自然拡散により培養液中に移動する精子の割合も格段に低くなる。一方、精子濃縮液中の運動精子は、自身の運動性により凹部４内の濃縮液中より底面２上の培養液中へと進出し、さらに底面２上を側壁１３へ向かって移動する。
30〜60分静置後、倒立顕微鏡または実体顕微鏡などを用い、底面２上に移動した運動精子のみを採取針などで採取する。このとき、滅菌された器具により培養液を部分的に遮断すると、培養液表面のミネラルオイルが遮断された部分に侵入して培養液を分割する。したがって、この操作により目的の運動精子の存在する培養液を部分的に分割して、より確実に該運動精子のみを採取することがかのうである。また、底面２上に凹部４の縁からの距離を示す目盛りが付けられていれば、精子の運動性の高低を明らかに認識でき、より好ましい。
【００１５】
図２に示すようにすり鉢状の第一凹部１４１が形成されたシャーレ１１を用いれば、第二凹部１４３に注入された精子懸濁液から培養液表面までの深さがより深くなるため、より運動性の高い運動精子のみが培養液表面へ移動することになる。したがって、より運動性の高い運動精子のみを採取することが可能となる。
【００１６】
【発明の効果】
本発明のシャーレは、底面に設けられた凹部に精子懸濁液を注入することにより、該懸濁液中に含まれる精子の培養液への自然拡散を抑制することができるため、運動精子のみを確実に採取することが可能となる。また、２段階で深くなるの凹部を設けることにより、本発明のシャーレはより運動性の高い精子のみを選択採取することが可能である。
【図面の簡単な説明】
【図１】 本発明の本発明のシャーレの一実施例を示す側面図および断面図である。
【図２】 本発明の本発明のシャーレの他の実施例を示す側面図および断面図である。
【符号の説明】
１、１１ シャーレ
２，１２ 底面
３、１３ 側壁
４ 凹部
１４１ 第一凹部
１４３ 第二凹部[0001]
[Technical field to which the invention belongs]
The present invention relates to a petri dish for selective collection of sperm used for selectively collecting motor sperm from a sperm suspension under a microscope.
[0002]
[Prior art]
Among fertility assistive technologies (ART), microinsemination (ICSI) for severe semen findings and other cases involves the whole process of sperm going up the female reproductive tract, fertilizing and fusing with eggs. It is a method to do in. According to this in vitro fertilization, even in cases where there are extremely few spermatozoa in the ejaculated semen or in cases where the sperm in the semen lacks the ability to invade the ovum by itself (fertilization ability) Fertilization is possible if there is a sperm and one egg. The sperm used here may be any of ejaculate, epididymis sperm or testicular sperm.
[0003]
Conventionally, in ART, it is important to use morphologically normal motor spermatozoa extracted from semen for fertilization. The motile sperm is concentrated in 1 to 5 ml of a density gradient body (hereinafter referred to as sperm suspension) containing sperm after concentration of semen and removal of seminal plasma by density gradient centrifugation using a density gradient body. It is obtained by a method (swim up method) in which the sperm is allowed to stand in the liquid and then swim up in the culture medium due to its own motility. However, there are many cases of poor semen findings that are subject to ICSI when the number of sperm in the semen is small or the motility rate of sperm is low, and it is not possible to recover sufficient motile sperm by such a swim-up method. .
[0004]
Therefore, for the collection of motor sperm in ICSI, a sperm suspension obtained by concentrating 1 to 10 ml of semen to 10 to 50 μl by density gradient centrifugation and a petri dish into which 50 to 100 μl of a culture solution has been introduced in advance are used. A method is used in which a sperm suspension is injected into the culture solution and then allowed to stand to recover the swollen moving sperm in the suspension. In this method, since a small amount of sperm suspension and culture solution are used, motile sperm can be collected under a microscope.
However, since the petri dish used in the motor sperm collection method in the ICSI has a smooth bottom surface, when the sperm suspension is injected into the culture solution introduced into the petri dish bottom surface, the suspension spreads over the whole culture solution by natural diffusion. Because it spreads, it is very difficult to separate motor and non-motor spermatozoa.
[0005]
[Problems to be solved by the invention]
In view of the above circumstances, an object of the present invention is to provide a petri dish that can suppress the influence of natural diffusion and can efficiently select and collect only motor sperm.
[0006]
[Means for Solving the Problems]
As a result of various intensive studies to solve the above problems, the present inventors have formed a recess having an internal volume capable of accommodating a concentrated sperm suspension for use in ICSI on the bottom of a conventional petri dish. Thus, it was found that a petri dish capable of collecting only motor spermatozoa without being affected by the natural diffusion of the suspension can be provided.
[0007]
That is, the present invention is a petri dish for selective collection of sperm having a bottom surface and a side wall, and having a recess having a volume of 0.01 to 0.2 ml on the bottom surface.
Further, the present invention is a petri dish for selective collection of sperm, which has a bottom surface and a side wall, the bottom surface has a shape that deepens stepwise, and the deepest portion is a recess having a volume of 0.01 to 0.2 ml. .
[0008]
[Action]
Since the petri dish of the present invention injects the sperm suspension into the recess provided on the bottom surface, it suppresses the natural dispersion of the suspension into the culture solution and advances from the recess due to its own mobility. It is easy to select and collect only motor sperm. Moreover, since the moving spermatozoa which advanced from the recessed part exists on the bottom surfaces other than the recessed part of the petri dish, it is easy to identify under a microscope, and the moving sperm can be collected efficiently.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the petri dish of the present invention will be described in detail based on preferred embodiments shown in the accompanying drawings, but the present invention is not limited to these descriptions.
FIG. 1 is a side view and a sectional view showing an embodiment of the petri dish of the present invention, and FIG. 2 is a side view and a sectional view showing another embodiment of the petri dish of the present invention.
[0010]
An embodiment of the present invention will be described with reference to FIG. The petri dish 1 shown in FIG. 1 is composed of a bottom surface 2 and a side wall 3, and preferably has a circular bottom surface 2, and a bottomed cylindrical body composed of a side wall 3 having a much smaller height than the diameter of the bottom surface 2. It is. The petri dish 1 contains a culture solution for maintaining sperm viability in advance, and a concentrated sperm-containing solution (a sperm suspension described later) is injected therein to select motor sperm to be used for ICSI. It will be collected. Accordingly, the petri dish 1 has an internal volume that allows the culture medium and sperm suspension to be accommodated, and preferred dimensions thereof are that the bottom surface 2 has a diameter of 30 to 120 mm, and the side wall 3 has a height. 3-10 mm. The lower the height of the side wall 3, the smaller the internal volume, but it becomes easier to perform the operation of injecting the sperm suspension and collecting the moving sperm.
The petri dish 1 is preferably made of a highly transparent material such as polystyrene, polyethylene, and polyamide that can be sterilized by electron beam.
[0011]
A concave portion 4 is provided on the bottom surface 2 of the petri dish 1. The concave portion 4 can accommodate a density gradient body containing sperm (referred to as a sperm suspension) concentrated by density gradient centrifugation using a density gradient body when selective collection of motor sperm for ICSI is performed. It has an internal volume. Specifically, the inner volume of the recess 4 is 0.01 to 0.2 ml, preferably 40 to 80 μl. The shape of the recess 4 is not particularly limited, but is preferably a bottomed cylinder having a diameter of 2 to 10 mm and a depth of 2 to 10 mm.
The concave portion 4 may be provided so as to be buried in the bottom surface 2 with a thickness on the bottom surface 2 of the petri dish 1, or may be provided so as to protrude downward from the bottom surface 2 as shown in FIG. . In the latter case, it is more preferable that the side wall 3 of the petri dish 1 forms a leg part 5 that projects downward from the bottom surface of the recess 4 because the petri dish 1 can be placed stably.
The concave portion 4 may be provided at any position on the bottom surface 2 of the petri dish 1, but is preferably provided at a substantially central portion of the bottom surface 2 in terms of molding the petri dish 1 and a sperm concentrate injection operation.
[0012]
Another embodiment of the present invention will be described with reference to FIG. The petri dish of the present invention may have a shape in which the bottom surface becomes deeper in steps, and is preferably a petri dish 11 having a bottom surface that becomes deeper in two steps as shown in FIG. The petri dish 11 is a bottomed cylindrical body composed of a bottom surface 12 and a side wall 13, preferably a circular bottom surface 12, and a side wall 13 having a much smaller height than the diameter of the bottom surface 12. The same thing as the petri dish 1 shown in FIG.
A first recess 141 is provided on the bottom surface 2 of the petri dish 11, and a second recess 143 is provided on the bottom surface 142 of the first recess 141. This 2nd recessed part 143 plays the same role as the recessed part 4 in the petri dish 1 shown in FIG. 1, and a preferable dimension and shape are prescribed | regulated similarly.
The 1st recessed part 141 provided in the bottom face 12 of the petri dish 11 is for selecting a sperm with higher mobility. The volume of the first concave portion 141 is smaller than the volume of the petri dish 11, and specifically, the diameter of the bottom surface 142 is 20 to 80 mm and the depth is 2 to 5 mm. The first recess 141 has a bottomed cylindrical shape, that is, the side wall 144 does not have to be perpendicular to the bottom surface 142, but rather the side wall 144 has a mortar shape having an angle with respect to the bottom surface 142. Preferred in operation.
[0013]
Further, the first concave portion 141 and the second concave portion 143 may be provided so as to be buried in the bottom surface 12 with a thickness on the bottom surface 12 of the petri dish 11, or downward from the bottom surface 12 as shown in FIG. It may be provided so that it may protrude. In the latter case, it is more preferable that the side wall 13 of the petri dish 11 forms a leg portion 15 that projects downward from the bottom surface 12.
The first concave portion 141 and the second concave portion 143 may be provided on any one of the bottom surfaces 12 of the petri dish 11. However, like the petri dish 1, the first concave portion 141 and the second concave portion 143 are preferably provided substantially at the center of the bottom surface 12. It is more preferable that the bottom surfaces of the first concave portion 141 and the second concave portion 143 are all concentric circles.
[0014]
Next, a method of using the petri dish of the present invention will be described using the petri dish 11 shown in FIG.
First, about 50 to 100 μl of a culture solution such as HTF (Human Tubal Fluid Medium) or Huecm (registered trademark, manufactured by Nipro Co., Ltd.) is introduced around the concave portion 4 of the petri dish 1, The culture surface is coated with mineral oil or the like. Next, 10-50 μm of sperm suspension concentrated by a known method such as density gradient centrifugation is injected into the recess 4 using a thin tubular instrument such as a syringe needle or glass capillar, Put.
Since the sperm suspension injected into the recess 4 has a much smaller contact area with the culture solution than when using a conventional petri dish, the proportion of sperm that moves into the culture solution by natural diffusion is also low. It will be much lower. On the other hand, the moving sperm in the sperm concentrate moves from the concentrated solution in the recess 4 to the culture solution on the bottom surface 2 due to its own mobility, and further moves on the bottom surface 2 toward the side wall 13.
After standing for 30 to 60 minutes, using an inverted microscope or a stereomicroscope, only the moving sperm that has moved to the bottom surface 2 is collected with a collection needle or the like. At this time, when the culture solution is partially blocked with a sterilized instrument, the culture solution is divided by entering the portion where the mineral oil on the surface of the culture solution is blocked. Therefore, it is possible to divide the culture solution containing the target motile spermatozoon partially by this operation and collect only the motile sperm more reliably. Moreover, if the scale which shows the distance from the edge of the recessed part 4 is attached on the bottom face 2, the motility of a sperm can be recognized clearly, and it is more preferable.
[0015]
If the petri dish 11 in which the mortar-shaped first concave portion 141 is formed as shown in FIG. 2, the depth from the sperm suspension injected into the second concave portion 143 to the surface of the culture solution becomes deeper. Only motile sperm with high mobility will move to the surface of the culture solution. Therefore, it is possible to collect only motor sperm with higher mobility.
[0016]
【The invention's effect】
Since the petri dish of the present invention can suppress the natural diffusion of sperm contained in the suspension into the culture solution by injecting the sperm suspension into the recess provided on the bottom surface, only the moving sperm Can be reliably collected. In addition, by providing a deep recess in two stages, the petri dish of the present invention can selectively collect only sperm with higher mobility.
[Brief description of the drawings]
FIG. 1 is a side view and a sectional view showing an embodiment of a petri dish of the present invention.
FIG. 2 is a side view and a sectional view showing another embodiment of the petri dish of the present invention.
[Explanation of symbols]
1, 11 Petri dish 2, 12 Bottom surface 3, 13 Side wall 4 Recess 141 First recess 143 Second recess

Claims (9)

底面と側壁とを有し、該底面に0.01〜0.2mlの容積を有する凹部が設けられてなる精子選択採取用シャーレ。A petri dish for selective collection of sperm, which has a bottom surface and side walls, and is provided with a recess having a volume of 0.01 to 0.2 ml on the bottom surface. 前記凹部は、シャーレの底面中央部に設けられ、底面の直径が2〜10mmであり、深さが2〜10mmである円柱形状を有する請求項１記載の精子選択採取用シャーレ。The petri dish for selective collection of spermatozoa according to claim 1, wherein the concave portion is provided at the center of the bottom surface of the petri dish, and has a cylindrical shape with a bottom surface diameter of 2 to 10 mm and a depth of 2 to 10 mm. 前記シャーレは、直径30〜120mmの円形底面と高さ3〜10mmの側壁とを有する、請求項１または２記載の精子選択採取用シャーレ。The petri dish for selective collection of sperm according to claim 1 or 2, wherein the petri dish has a circular bottom surface with a diameter of 30 to 120 mm and a side wall with a height of 3 to 10 mm. 前記シャーレの底面には、前記凹部の縁からの距離を示す目盛りが付けられてなる、請求項１〜３のいずれかに記載の精子選択採取用シャーレ。The petri dish for selective collection of spermatozoa according to any one of claims 1 to 3, wherein a scale indicating a distance from an edge of the concave portion is attached to a bottom surface of the petri dish. 底面と側壁とを有し、該底面が段階的に深くなる形状を有し、その最深部が0.01〜0.2mlである容積を有する凹部である、請求項１記載の精子選択採取用シャーレ。The petri dish for selective collection of spermatozoa according to claim 1, wherein the petri dish has a bottom surface and a side wall, the bottom surface has a shape that deepens stepwise, and the deepest portion has a volume of 0.01 to 0.2 ml. 底面と側壁とを有し、該底面に第一凹部が設けられ、該第一凹部の底面に0.01〜0.2mlの容積を有する第二凹部が設けられてなる、請求項５記載の精子選択採取用シャーレ。The sperm selective collection according to claim 5, comprising a bottom surface and a side wall, wherein the bottom surface is provided with a first recess, and the bottom surface of the first recess is provided with a second recess having a volume of 0.01 to 0.2 ml. Petri dish for. 前記第一凹部は、シャーレの底面中央部に設けられ、底面の直径が20〜80mmであり、深さが2〜5mmであり、前記第二凹部は、該第一凹部の底面中央部に設けられ、底面の直径が2〜10mmであり、深さが2〜10mmである円柱形状を有する、請求項６記載の精子選択採取用シャーレ。The first recess is provided at the center of the bottom of the petri dish, the diameter of the bottom is 20 to 80 mm, the depth is 2 to 5 mm, and the second recess is provided at the center of the bottom of the first recess. The petri dish for selective collection of spermatozoa according to claim 6, wherein the petri dish has a cylindrical shape with a bottom surface diameter of 2 to 10 mm and a depth of 2 to 10 mm. 前記シャーレは、直径30〜120mmの円形底面と高さ3〜10mmの側壁とからなる、請求項６または７記載の精子選択採取用シャーレ。The petri dish for selective collection of sperm according to claim 6 or 7, wherein the petri dish is composed of a circular bottom having a diameter of 30 to 120 mm and a side wall having a height of 3 to 10 mm. 前記第一凹部の底面には、前記第二凹部の縁からの距離を示す目盛りが付けられてなる、請求項６〜８のいずれかに記載の精子選択採取用シャーレ。The petri dish for selective collection of sperm according to any one of claims 6 to 8, wherein a scale indicating a distance from an edge of the second concave portion is attached to a bottom surface of the first concave portion.
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