Live cells imaging

Yano Y, Matsuzaki K (2009) Tag-probe labeling methods for live-cell imaging of membrane proteins. Biochim Biophys Acta-Biomembranes 1788 2124—2131... 

Wang M, Gao M, Miller KD et al (2009) Simple synthesis of carbon-11 labeled styryl dyes as new potential PET RNA-specific, living cell imaging probes. Eur J Med Chem 44 2300-2306... 

The potential of live cell imaging to address mechanisms of cellular biology is ever expanding. Directed protein-tagging techniques have been used to visualize nascent versus mature protein in vivo (Rodriguez et al., 2006). This technique involves the use of arsenic-based dyes, such as FiAsH or ReAsH, which bind to tetracysteine (TC) tags (Zhang et al, 2002). In addition, photo-activatable variants of GFP have been shown to determine the kinetics of protein movement in live cells (Patterson and Lippincott-Schwarz, 2002). Furthermore, techniques such as FRET and the... 

Yudushkin, I. A., Schleifenbaum, A., Kinkhabwala, A., Neel, B. G., Schultz, C. and Bastiaens, P. I. H. (2007). Live-cell Imaging of enzyme-substrate interaction reveals spatial regulation of PTP1B. Science 315, 115-119. 

Brown, A. Live-cell imaging of slow axonal transport in cultured neurons. Methods Cell Biol. 71 305-323,2003. 

Niino Y, Hotta K, OkA K (2009) Simultaneous live cell imaging using dual FRET sensors with a single excitation light. PLoS One 4 e6036... 

LysoTracker and LysoSensor probes Several commercially available weakly basic amines can be used to stain lysosomes. They selectively accumulate in acidic organelles when applied in very low concentrations (50 nM) and directly before imaging. For live cell imaging, keep in mind that lysosomal probes can exhibit an alkalinizing effect on the lysosomes, such that longer incubation time can induce an increase in lysosomal pH (129). 

Microtubules are used to cover relatively long distances in the cell. Therefore, the short distance between the cell surface and the early endosomes does not require microtubules (94,136). They are involved in the later steps in endocytosis early endosomes accumulate endocytic material for about 10 minutes and then generate transport vesicles (0.5 pm in diameter), which will be taken to the cell center with a relatively high traveling speed of Ipm/sec (with velocities of up to 2.5pm/sec) (95,137). These transport vesicle move on microtubule-tracks to the cell center (and to the ly sosome) (94). When performing live cell imaging studies, it should be kept in mind that microtubules are extremely sensitive to ultraviolet light, which causes their polymerization. 

The potential of live cell imaging techniques, which overcome the drawbacks of in vitro techniques, is just beginning to be realized. A major advance that is... 

Cell based assay, Live cell imaging... 

The outline of this chapter is as follows. First, we discuss the methods of THG microscopy and CARS microspectroscopy and outline the major developments over the past years, emphasizing the application aspect of this work. Then, we discuss the application of these spectroscopy tools for several microfluidic problems, such as live-cell imaging and protein crystallization. 

Stephens, D.J., and VJ. Allan. 2003. Light microscopy techniques for live cell imaging. Science 300 82-86. 

Beatty KE, Fisk JD, Smart BP et al (2010) Live-cell imaging of cellular proteins by a strain-promoted azide-alkyne cycloaddition. Chembiochem 11 2092-2095... 

Robert D. Goldman and David L. Spector (Editors). Live Cell Imaging. A Laboratory Manual. 2005. 

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