Microinjection living cells

Need for micromanipulatory procedure. The metabolic studies require the observation of dynamic changes triggered by microinjection of metabolites. The usefulness of both spectral and topographic operation is critically dependent upon the ability to micromanipulate (microinject) living cells, in order to measure changes in injected cells and their neighbors. 

Many derivatives of fluorescein containing a reactive group at the C-5 position are commercially available [11], Fluorescein isothiocyanate, for example, is widely used as protein tag [12]. These substances have essentially the same spectroscopic properties as the parent compound with the additional capability of binding covalently to proteins. Because of their high emission quantum yields, fluorescein conjugates are extensively used as tracers for microinjection in living cells to gather information on the structure and function of cells, localization of proteins, and cell-to-cell and intracellular diffusion [13-17]. 

Mangeat, P. H., and Burridge, K. (1984). Immunoprecipitation of nonerythrocyte spectrin within live cells following microinjection of specific antibodies Relation to cytoskeletal structures./ Cell. Biol. 98, 1363-1377. 

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. 

II. Microinjection of Laniin Antibodies into Living Cells... 

Ansorge, W. (1982). Improved system for capillary microinjection into living cells. Exp. Cell Res. 140,31-37. 

Microinjection is a process in which very fine needles are used to inject substances into a single living cell under the microscope. Small needles of roughly 0.5-5 pm in diameter are used to inject the desired contents into the desired subcellular compartment. 

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