Piezo-actuated mouse intracytoplasmic sperm injection (ICSI)

被引：121

作者：

Yoshida, Naoko ^{[1
]}

Perry, Anthony C. F. ^{[1
]}

机构：

[1] RIKEN, Ctr Dev Biol, Lab Mammalian Mol Embryol, Chuo Ku, Kobe, Hyogo 6500047, Japan

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 02期

关键词：

D O I：

10.1038/nprot.2007.7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The mouse is a genetically tractable model organism widely used to study mammalian development and disease. However, mouse metaphase II (mII) oocytes are exquisitely sensitive and intracytoplasmic sperm injection (ICSI) with conventional pipettes generally kills them. This problem can be solved with piezo-actuated micromanipulation, in which the piezo-electric effect (crystal deformation in response to an externally applied voltage) propels a microinjection needle tip forward in a precise and rapid movement. Piezo-actuated micromanipulation enhances the penetration of membranes and matrices, and mouse ICSI is a major application. Here we describe a comprehensive, step-by-step mouse piezo ICSI protocol for non-specialists that can be completed in 2 - 4 h. The protocol is a basic prelude to multiple applications, including nuclear transfer cloning, spermatid injection, blastocyst injection, mII transgenesis, and streamlining micromanipulation in primates and livestock. Moreover, piezo ICSI can be used to obtain offspring from 'dead' (non-motile) sperm, enabling trivial sperm freezing protocols for mouse strain storage and shipment.

引用

页码：296 / 304

页数：9

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