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Lakowicz’s classic book 17 is highly recommended for acquiring deep understanding on fluorescence spectroscopy. Tsien in 2008.Ī large number of fluorescence imaging techniques have been developed. This break-through resulted in the awarding of the Noble Prize in Chemistry to Osamu Shimomura, Martin Chalfie, and Roger Y. 16 The high specificity of fluorescence microscopy is in part the result of the discovery and subsequent cloning of the green fluorescent protein (GFP) and its many variants, which made it possible to express a fluorescent label fused to a protein of interest throughout a given cell or organism. Among other purposes, fluorescence microscopy can be used to measure properties of molecular and cellular movements 13- 14 the cellular location of biomolecules 15 and interactions between biomolecules. Finally, we provide a brief, yet rather complete, summary of single cell manipulation techniques.ĭue to its high sensitivity and specificity, florescence microscopy is commonly used for a broad range of applications in cell biology. Particular attention is paid to the emerging orientation and rotational tracking of single probes linked to mechanistic functions and differentiated structures of biological interest. Next we emphasize specific imaging experiments that highlight the types of findings that are possible at the nexus of microscopy, nanoprobes, and live cells. As such, much of our emphasis in the first several sections of this paper is on imaging platforms, with a focus on design details that are important to single cell imaging experiments. 6- 12 This review is designed to give an overview of the tools that are being specifically used to accomplish single cell imaging.
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2- 7 Likewise, exceptional review papers that have discussed the full spectrum of nanoparticle probes and their properties have appeared recently.
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Reviews with a distinctly biological flavor have been published recently, and these alternative reviews focus on specific details of the cell and the processes that occur within. It is not our intent to provide a comprehensive review of the types of experiments or the areas of cell research that are ongoing. The purpose of this review is to highlight the key advances that have occurred in the past several years in the field of single cell optical imaging. Advances in laser, camera, and imaging processing technologies have also played a crucial role in the burgeoning field of single cell imaging, because they have brought into view the fast processes that would normally escape the human eye. Enter a variety of molecular and nanoparticle probes that are capable of tagging and pinpointing the location of biological components that would otherwise be invisible under the microscope. Size, speed, sensitivity, and additional concerns plague the microscopist who wants to peek inside of a cell. Seeing is not always a possibility in biological systems. Ultimately, scientists want to ‘see to believe’ when it comes to an explanation of the complex inner workings of cells, but therein lies a complication. The cell is a basic yet complicated ‘unit’ of interest to biology, just as the atom is to chemists. 1 In the three and a half centuries since Hooke’s day, both the microscope and our understanding of the cell have been vastly improved upon, and the current outlook suggests that the symbiotic relationship between the microscope and the cell will continue to flourish into the foreseeable future. In his 1665 treatise, Micrographia, Robert Hooke described the many observations he had made using a microscope, including compartment-like structures in cork samples that he termed ‘cells’.