Gene Delivery by Direct Injection (Microinjection) Using a Pulsed-Flow System
This protocol was adapted from “Gene Delivery by Direct Injection and Facilitation of Expression by Mechanical Stretch,” Chapter 4, in Live Cell Imaging (eds. Goldman and Spector). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.INTRODUCTION
This protocol describes a method for pulsed-flow microinjection using the Eppendorf FemtoJet injector and Eppendorf InjectMan; this is the most common type of pulsed-flow microinjection system currently being used. The advantage of this type of system over a controlled-flow system is that much more control is available over the injection parameters, reducing variability in injections. In addition, the system allows a diagonal insertion of the needle into the cell, causing a direct piercing of the cell. This is fast and may do less damage to the cell than a traditional system, in which the cell is briefly distorted before it is pierced.
