请求 精子如何计数

yangpan521

    急求 望高人指点

淡定

用24孔板，每孔加入约500ul的双蒸水，放在摇床上，慢速摇动，将附睾头剪取约0.03克左右的组织放入24孔板中，同时附睾要称重，直至将标本收集完成，建议不要一次杀死太多的老鼠，时间越长在24孔板的精子的活性就越弱了，有可能死亡的。直接用200ul的枪头吸取20ul的混悬液，用计数板计数，按照血细胞计数操作即可。以上意见仅供参考。

aulenuyah

与楼上说的差不太多，不过稀释精液的不能用双蒸水吧，最好用PBS之类的缓冲体系，混匀后用甲醛等物质杀死精子，然后再用血细胞计数板计数

ldy689689

可以做流式技术吧
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279042256

生理盐水稀释后加甲醛杀死后上细胞计数板慢慢数吧，

liuyul03

精子计数其实比较简单，可以浓度较高的盐水即可，如3%，然后上计数板，根据数左不数右，数上不数下的原则计数即可。个人浅见。

精神病院小萝卜

用casa仪呀

mansnoopy

回复 yangpan521 的帖子
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6 B; n' c7 c& H1 N9 [9 H8 ~4 V, ]Cauda epididymal sperm counts For sperm count analyses, each cauda epididymis was placed in a clean 1.5 mL microcentrifuge tube containing 500 lL distilled water and minced using a sterile Squisher_ (Zymo Research Corp., CA, USA). The tube was vortexed briefly and a 100 lL aliquot of the resulting suspension was placed in an IDENT-stain tube (Hamilton-Thorne Biosciences) with 100 lL distilled water. The tube was again vortexed and incubated for 3 min at room temperature. A 5 lL aliquot was placed on a 20 lm chamber slide (X-CEL; Hamilton-Thorne Biosciences), and spermatoazoa were counted using the IDENT option of the Oval Head Toxicology programme (Hamilton-Thorne Biosciences). A minimum of 40 scans per individual were recorded.

gdeizhangxiujua

用3%KCL溶液，按比例稀释精子，10min后，用普通计数板计数，很简单的。

daixiuliang

CASA检测，按上述方法，取10ul 精子悬液，上机检测。到底是取附睾头还是尾？