Benefits

• Increase embryoviabilityfor theinjected embryos
• Flexible system
• Integrates with WPI’s PV820/PV830, as well as other popular microinjection systems, like the Eppendorf Femtojet® and the Narishige injectors (Femtojet® 4I is a registered Trademark of Eppendorf.)
• Microinjection of diverse compounds and biomolecules – DNA, RNA, proteins
• Pre- and post-implantation in embryos of various species – mice, rodents, monkeys, bovine, pigs, zebrafish, etc.
• Easy to operate cell injector– Hands-free operation with foot switch control
• User adjustable frequency and voltage – four programmable protocols
• Small, portable unit saves precious benchtop space
• Works with all major inverted microscopes

Target Applications

• Cell microinjection into oocytes and pre-implantation stage mammalian embryos, including microinjection of CRISPR-Cas9 reagents into the cytoplasm of two-cell stage embryos
• Pronuclear rodent zygote microinjection
• Gene silencing in zebrafish

Simple, Elegant Solution

The new WPI MICRO-ePORE™ pinpoint cell penetrator is a simple and versatile system that can be used for efficient cellmicroinjection of a diverse array of compounds and biomolecules into oocytes and pre-implantation stage mammalian embryos. Patent pending Flutter Electrode Technology assists in small, clean, precise membrane penetration without tearing or damaging the membrane when you are working on the microinjection of transgenic animals or cell manipulation.

Pinpoint Cell Penetrator vs. Electroporation

Electroporation uses a pulse of electricity in a mediumto open pores in the cellmembranes of cells in the electroporation cuvette in order to penetrate the cell membrane and introduce genetic material.

WPI"s MICRO-ePORE™PinpointCellPenetratoroffers several advantages over traditional electroporation for the purpose of cellmicroinjection.

• Our pinpoint cellmicroinjectorusesamuch lowervoltage to open a port into the cell membrane.
• Electroporation is a shotgun approach that opens many pores in the cell membrane. In contrast, our pinpoint cell microinjector targets a specific area of the cell membrane at the very point of cellular microinjection.
• Embryo survivabilityis significantly higher when using pinpoint cell penetration versus electroporation.

Experimental Data

Electrophysiological systems utilizing negative capacitance have been routinely used for cellmicroinjection of a variety of biomolecules into mammalian oocytes, as well as pre-implantation and post-implantation embryos in developmental biology studies. The system which is no longer available, the intracellular amplifier WPI Cyto721, allows the needle to pierce the cell membrane with minimal physical trauma. More recently this technique has been applied to genetic transfer in the microinjection of CRISPR/Cas9 reagents into two-cell stage mouse embryos.The authors demonstrated significant increase in the knock-in efficiency and high viability of embryos using their method.

The new MICRO-ePORE™ pinpoint cell penetrator offers a unique solution for cellmicroinjection resulting in high embryoviability. The instrument creates an oscillating electric field at a localized site on the membrane immediately beneath the site of injection. The MICRO-ePORE™ creates small, reversible holes in the plasma membrane through which material is microinjected. The researcher determines the amplitude and frequency of the signal that best suits the application. In contrast to conventional cellmicroinjection, in targeted microinjection using the MICRO-ePORE™, the membrane does not tear and thus allows for superior embryoviability. The technique is simple and elegant. The new MICRO-ePORE™ cell penetrator prototype has been successfully tested in mouse and primate pre-implantation embryos, as well as gene silencing in zebrafish tails.

MICRO-ePORE™ was designed for a range of applications including generation of CRISPR/Cas9 mediated knock-in mice with large insertions by microinjection into two-cell stage embryos with high viability.7 The MICRO-ePORE™ has delivered accurate microinjection of morpholino oligomers (anti-sense “knockdown”) in zebrafish tails.

Well-Style Reference Electrode Option

Probe-Style Reference Electrode Option

This is showing the probe-style reference electrode with an Eppendorf injector, by Fangtao Chi of UCLA/MCDB and Utpal Banerjee.

Setting up Your MICRO-ePORE™

The new WPI MICRO-ePORE™ Pinpoint Cell Penetrator is a simple and versatile system that can be used for efficient microinjection of a diverse array of compounds and biomolecules into oocytes and pre-implantation stage mammalian embryos. Patent pending Flutter Electrode Technology assists in small, clean, precise membrane penetration without tearing or damaging the membrane. Here Gabe sets up the system and connects all the components.