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Sample Preparation for Microinjection

General Comments
Microinjection is the loading or transfer of a dissolved substance into a living cell. The microscopic tip of the glass microcapillary has an inner diameter between 0.2 and 1 m [1]. This capillary is back loaded with the substance to be transferred into the cells cultured for microinjection. Typical substances include purified antibodies, DNA, RNA, peptides, or oligonucleotides. To visualize and evaluate the success of a microinjection experiment these substances are typically mixed with dyes or labeled with fluorescent markers such as flourescein or rhodamine. After the capillary has pierced the cell, a certain amount of transfer substance (approximately 10% of the cell volume) is transferred from the capillary into the cell due to pressure exerted on the capillary via the microinjector [2]. Because the diameter of the capillary is very small, particles in the injection solution can quickly result in blockage of the capillary. Therefore, exchange of the capillary is a common practice during microinjection experiments and the frequency of the capillary exchange depends on the concentration and clarity of substance injected. To maximize the number of injections which can be performed with a single capillary, the injection substance should be centrifuged for 1015 minutes at a minimum of 10,000 x g before back loading the capillary. As the cross section of the capillary tip is small and very little liquid would be found
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