Fluorescent reporters and methods to analyze fluorescent signals.

The use of fluorescent reporters and the development of new imaging technologies have revolutionized studies in cell biology. During recent years the number of fluorescent proteins offering the ability to visualize the distribution of proteins, organelles, and cells has increased tremendously. In parallel, the imaging tools available were refined rapidly enabling now the use of a huge spectrum of specialized methods to explore the cellular and subcellular localization and dynamics of fluorescently tagged markers. This chapter presents an overview of fluorescent reporters and methods available, and describes a selection of those that are routinely applicable in imaging studies using Dictyostelium discoideum.

[1]  R. Tsien,et al.  Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein , 2004, Nature Biotechnology.

[2]  David W Piston,et al.  Fluorescent proteins at a glance , 2011, Journal of Cell Science.

[3]  Dynamics of the Dictyostelium cytoskeleton during chemotaxis. , 1992, The New biologist.

[4]  W. Baumeister,et al.  Dynamic organization of the actin system in the motile cells of Dictyostelium , 2004, Journal of Muscle Research & Cell Motility.

[5]  G. Whitesides,et al.  Generation of Solution and Surface Gradients Using Microfluidic Systems , 2000 .

[6]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

[7]  G. Gerisch,et al.  Coronin involved in phagocytosis: Dynamics of particle-induced relocalization visualized by a green fluorescent protein tag , 1995, Cell.

[8]  Adriaan B Houtsmuller,et al.  Fluorescence recovery after photobleaching: application to nuclear proteins. , 2005, Advances in biochemical engineering/biotechnology.

[9]  T. Holak,et al.  Lifeact: a versatile marker to visualize F-actin , 2008, Nature Methods.

[10]  G. Gerisch,et al.  Myristoylated and non‐myristoylated forms of the pH sensor protein hisactophilin II: intracellular shuttling to plasma membrane and nucleus monitored in real time by a fusion with green fluorescent protein. , 1996, The EMBO journal.

[11]  V. Verkhusha,et al.  Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light , 2006, Nature Biotechnology.

[12]  B. Giepmans,et al.  Immunolabeling artifacts and the need for live-cell imaging , 2012, Nature Methods.

[13]  R. Tsien,et al.  A monomeric red fluorescent protein , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Markus Fischer,et al.  A brilliant monomeric red fluorescent protein to visualize cytoskeleton dynamics in Dictyostelium , 2004, FEBS letters.

[15]  James B. Pawley,et al.  Fundamental Limits in Confocal Microscopy , 2006 .

[16]  L. Trusolino,et al.  Scatter-factor and semaphorin receptors: cell signalling for invasive growth , 2002, Nature Reviews Cancer.

[17]  Maja A. Marinović,et al.  A dual role for Rac1 GTPases in the regulation of cell motility , 2011, Journal of Cell Science.

[18]  C. Janetopoulos,et al.  FRAP analysis of chemosensory components of Dictyostelium. , 2009, Methods in molecular biology.

[19]  S. J. Wright,et al.  Introduction to confocal microscopy. , 2002, Methods in cell biology.

[20]  I. Charo,et al.  Chemokines in the pathogenesis of vascular disease. , 2004, Circulation research.

[21]  T. Funatsu,et al.  The dynamics of pre-mRNAs and poly(A)+ RNA at speckles in living cells revealed by iFRAP studies. , 2008, Experimental cell research.

[22]  D Zicha,et al.  Fluorescence localization after photobleaching (FLAP): a new method for studying protein dynamics in living cells , 2002, Journal of microscopy.

[23]  C. Janetopoulos,et al.  Dynamic localization of G proteins in Dictyostelium discoideum , 2009, Journal of Cell Science.

[24]  Kurt I. Anderson,et al.  Recent advances using green and red fluorescent protein variants , 2007, Applied Microbiology and Biotechnology.

[25]  J. Spudich,et al.  Myosin dynamics in live Dictyostelium cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[26]  P. V. van Haastert,et al.  A new set of small, extrachromosomal expression vectors for Dictyostelium discoideum. , 2009, Plasmid.

[27]  T. Bretschneider,et al.  Differential localization of the Dictyostelium kinase DPAKa during cytokinesis and cell migration , 2004, Journal of Muscle Research & Cell Motility.

[28]  Richard N. Day,et al.  Fluorescent protein spectra. , 2001, Journal of cell science.

[29]  A. Nagasaki,et al.  Multiple myosin II heavy chain kinases: roles in filament assembly control and proper cytokinesis in Dictyostelium. , 2005, Molecular biology of the cell.

[30]  S. Lukyanov,et al.  Tracking intracellular protein movements using photoswitchable fluorescent proteins PS-CFP2 and Dendra2 , 2007, Nature Protocols.

[31]  A. Müller-Taubenberger Application of fluorescent protein tags as reporters in live-cell imaging studies. , 2006, Methods in molecular biology.

[32]  T. Misteli,et al.  A Kinetic Framework for a Mammalian RNA Polymerase in Vivo , 2002, Science.

[33]  T. Terwilliger,et al.  Engineering and characterization of a superfolder green fluorescent protein , 2006, Nature Biotechnology.

[34]  J. Rädler,et al.  Chemotactic cell trapping in controlled alternating gradient fields , 2011, Proceedings of the National Academy of Sciences.

[35]  S. Diez,et al.  Dynamic Actin Patterns and Arp2/3 Assembly at the Substrate-Attached Surface of Motile Cells , 2004, Current Biology.

[36]  P. Devreotes,et al.  Receptor-Mediated Activation of Heterotrimeric G-Proteins in Living Cells , 2001, Science.

[37]  S. Yumura,et al.  A novel mitosis-specific dynamic actin structure in Dictyostelium cells , 2007, Journal of Cell Science.

[38]  A. Miyawaki,et al.  An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[39]  C. Beta Spatiotemporal stimulation of single cells using flow photolysis. , 2009, Methods in molecular biology.

[40]  E. Bodenschatz,et al.  Using microfluidic channel networks to generate gradients for studying cell migration. , 2005, Methods in molecular biology.

[41]  D. Weitz,et al.  Mechanical strain in actin networks regulates FilGAP and integrin binding to Filamin A , 2011, Nature.

[42]  D. Wiersma,et al.  Reduced protein diffusion rate by cytoskeleton in vegetative and polarized dictyostelium cells. , 2001, Biophysical journal.

[43]  W. Rappel,et al.  Dictyostelium discoideum chemotaxis: threshold for directed motion. , 2006, European journal of cell biology.

[44]  Luke P. Lee,et al.  Microfluidics-based systems biology. , 2006, Molecular bioSystems.

[45]  Cornelis J. Weijer,et al.  Paxillin is required for cell-substrate adhesion, cell sorting and slug migration during Dictyostelium development , 2005, Journal of Cell Science.

[46]  M. Dubin,et al.  A versatile set of tagged expression vectors to monitor protein localisation and function in Dictyostelium. , 2010, Gene.

[47]  K. Anderson,et al.  Fluorescence lifetime imaging: association of cortical actin with a PIP3-rich membrane compartment. , 2008, European journal of cell biology.

[48]  Annette Müller-Taubenberger,et al.  A Lim protein involved in the progression of cytokinesis and regulation of the mitotic spindle. , 2003, Cell motility and the cytoskeleton.

[49]  T. Jin,et al.  Using quantitative fluorescence microscopy and FRET imaging to measure spatiotemporal signaling events in single living cells. , 2006, Methods in molecular biology.

[50]  R. Tsien,et al.  Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer , 1996, Current Biology.

[51]  J. Wiedenmann,et al.  EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[52]  C. Beta,et al.  Microfluidic tools for quantitative studies of eukaryotic chemotaxis. , 2011, European journal of cell biology.

[53]  Eberhard Bodenschatz,et al.  Flow photolysis for spatiotemporal stimulation of single cells. , 2007, Analytical chemistry.

[54]  S. Paddock Principles and practices of laser scanning confocal microscopy , 2000, Molecular biotechnology.

[55]  S. Lukyanov,et al.  Fluorescent proteins and their applications in imaging living cells and tissues. , 2010, Physiological reviews.

[56]  Monica L. Skoge,et al.  Gradient sensing in defined chemotactic fields. , 2010, Integrative biology : quantitative biosciences from nano to macro.

[57]  R. Tsien,et al.  Evolution of new nonantibody proteins via iterative somatic hypermutation. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[58]  Jeffrey G. Williams,et al.  The Dictyostelium prestalk cell inducer DIF regulates nuclear accumulation of a STAT protein by controlling its rate of export from the nucleus , 2003, Development.

[59]  D. Eccles Hereditary cancer: guidelines in clinical practice. Breast and ovarian cancer genetics. , 2004, Annals of oncology : official journal of the European Society for Medical Oncology.

[60]  George H. Patterson,et al.  A Photoactivatable GFP for Selective Photolabeling of Proteins and Cells , 2002, Science.

[61]  C. Beta,et al.  Flow-induced control of chemical turbulence. , 2011, The Journal of chemical physics.

[62]  S. Yumura Myosin II dynamics and cortical flow during contractile ring formation in Dictyostelium cells , 2001, The Journal of cell biology.

[63]  L. Eichinger,et al.  A Coronin7 Homolog with Functions in Actin-driven Processes* , 2010, The Journal of Biological Chemistry.

[64]  L. Eichinger,et al.  A Cytohesin Homolog in Dictyostelium Amoebae , 2010, PloS one.

[65]  Joachim Goedhart,et al.  Bright monomeric red fluorescent protein with an extended fluorescence lifetime , 2007, Nature Methods.

[66]  G. Gerisch Imaging actin cytoskeleton dynamics in Dictyostelium chemotaxis. , 2009, Methods in molecular biology.

[67]  Till Bretschneider,et al.  Subsecond reorganization of the actin network in cell motility and chemotaxis. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[68]  G. Drummen,et al.  Advanced Fluorescence Microscopy Techniques—FRAP, FLIP, FLAP, FRET and FLIM , 2012, Molecules.

[69]  M. Titus,et al.  Talin influences the dynamics of the myosin VII-membrane interaction. , 2007, Molecular biology of the cell.