Piezo-actuated mouse intracytoplasmic sperm injection (ICSI)

Research output: Contribution to journalArticle

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Abstract

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.
Original languageEnglish
Pages (from-to)296-304
Number of pages9
JournalNature Protocols
Volume2
Issue number2
DOIs
Publication statusPublished - 2007

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Intracytoplasmic Sperm Injections
Micromanipulation
Piezoelectricity
Cloning
Freezing
Needles
Agriculture
Metaphase
Spermatozoa
Membranes
Crystals
Electric potential
Gene Transfer Techniques
Injections
Spermatids
Microinjections
Livestock
Blastocyst
Primates
Oocytes

Cite this

Piezo-actuated mouse intracytoplasmic sperm injection (ICSI). / Yoshida, N; Perry, Anthony C F.

In: Nature Protocols, Vol. 2, No. 2, 2007, p. 296-304.

Research output: Contribution to journalArticle

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