Microinjection methods

Example ofSymplastic Tracing Using Microinjection Methods ... 

Import of aminoacyl-tRNA into living cells is another approach toward in vivo production of nonnatural mutant proteins. Dougherty and coworkers microinjected [41] or electroporated [44] an aminoacyl-tRNA/mRNA pair into Xenopus oocyte to synthesize fluorescently labeled acetylcholine receptor. The microinjection method is applicable to any type of tRNA and amino acid, but the number of cells that can be treated at one time is very limited. 

Recent studies utilizing microinjection of a neurotoxin to induce chemical lesions have implicated the pons (PoUacek et al. 2004), raphe nuclei (Jakus et al. 1998), and lateral tegmental field (Jakus et al. 2000) in the production of coughing. The microinjection method is best suited to the study of regions or nuclei of the brain... 

Millar J, Simultaneous in vivo voltammetric and electrophysiological recording with carbon fiber microelectrodes, Conn PM, ed., Electrophysiology and microinjection. Methods in Neurosciences, Academic Press, San Diego, 4, 143-154, 1991. 

Blinks, J. R., et al. (1978). Practical aspects of the use of aequorin as a calcium indicator Assay, preparation, microinjection, and interpretation of signals. Method. Enzymol. 57 292-328. 

Classical gene transfer methods still in use today are diethylamino ethyl (DEAE)-dextran and calcium phosphate precipitation, electroporation, and microinjection. Introduced in 1965, DEAE-dextran transfection is one of the oldest gene transfer techniques [2]. It is based on the interaction of positive charges on the DEAE-dextran molecule with the negatively charged backbone of nucleic acids. The DNA-DEAE-dextran complexes appear to adsorb onto cell surfaces and be taken up by endocytosis. 

Frothingham EP, Basbaum Al. 1992. Construction of a microdialysis probe with attached microinjection catheter. J Neu-rosci Methods 43(2-3) 181-188. 

Recently, a method has been developed to microinject cell-polymer suspensions into 3-D collagen matrices in a 96-well plate format. This was used to perform a proof-of-principle screen of known signaling pathway inhibitors in murine breast cancer cells. The cells were visualized by staining of the actin cytoskeleton and effects of the compounds measured using multiparametric imaging. The system also proved amenable to the use of enzymatically... 

Reid SW, Tessarollo L (2009) Isolation, microinjection and transfer of mouse blastocysts. Methods Mol Biol 530 269-285... 

Traditionally protein-protein interactions studies have been performed in vitro after isolation and purification of individual proteins. While some in vivo or in situ protein-protein interaction studies can be performed by traditional methods using microinjection of purified proteins into oocytes, technical complexities limit the number of proteins that can be studied. Furthermore, many putative proteins of interest, predicted by genomic analysis, are not characterized and cannot be used in such studies. Some of the limitations posed by traditional methods have been overcome by use of yeast two-hybrid systems. These systems allow studies of many recombinant test proteins... 

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