Micromirror structured illumination microscope for high-speed in vivo drosophila brain imaging.

Genetic tools and especially genetically encoded fluorescent reporters have given a special place to optical microscopy in drosophila neurobiology research. In order to monitor neural networks activity, high speed and sensitive techniques, with high spatial resolution are required. Structured illumination microscopies are wide-field approaches with optical sectioning ability. Despite the large progress made with the introduction of the HiLo principle, they did not meet the criteria of speed and/or spatial resolution for drosophila brain imaging. We report on a new implementation that took advantage of micromirror matrix technology to structure the illumination. Thus, we showed that the developed instrument exhibits a spatial resolution close to that of confocal microscopy but it can record physiological responses with a speed improved by more than an order a magnitude.

[1]  G. S. Kino,et al.  Real‐time confocal scanning optical microscope , 1988 .

[2]  Stephan J. Sigrist,et al.  Presynapses in Kenyon Cell Dendrites in the Mushroom Body Calyx of Drosophila , 2011, The Journal of Neuroscience.

[3]  Thomas Preat,et al.  PKA Dynamics in a Drosophila Learning Center: Coincidence Detection by Rutabaga Adenylyl Cyclase and Spatial Regulation by Dunce Phosphodiesterase , 2010, Neuron.

[4]  A. Fiala,et al.  Genetically Expressed Cameleon in Drosophila melanogaster Is Used to Visualize Olfactory Information in Projection Neurons , 2002, Current Biology.

[5]  P. Saggau,et al.  High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators. , 1997, Biophysical journal.

[6]  Ruikang K. Wang,et al.  Penetration kinetics of dimethyl sulphoxide and glycerol in dynamic optical clearing of porcine skin tissue in vitro studied by Fourier transform infrared spectroscopic imaging. , 2008, Journal of biomedical optics.

[7]  H. Bellen,et al.  Transgenesis upgrades for Drosophila melanogaster , 2007, Development.

[8]  Ronald L. Davis,et al.  Dynamics of Learning-Related cAMP Signaling and Stimulus Integration in the Drosophila Olfactory Pathway , 2009, Neuron.

[9]  K. Fujita.  [Two-photon laser scanning fluorescence microscopy]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[10]  Jerome Mertz,et al.  Wide-field fluorescence sectioning with hybrid speckle and uniform-illumination microscopy. , 2008, Optics letters.

[11]  Jerome Mertz,et al.  Fast optically sectioned fluorescence HiLo endomicroscopy. , 2012, Journal of biomedical optics.

[12]  J. Zerubia,et al.  Gaussian approximations of fluorescence microscope point-spread function models. , 2007, Applied optics.

[13]  Gero Miesenböck,et al.  Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins , 1998, Nature.

[14]  R. Webb.  Confocal optical microscopy , 1996 .

[15]  Ronald L. Davis,et al.  Drosophila alpha/beta mushroom body neurons form a branch-specific, long-term cellular memory trace after spaced olfactory conditioning. , 2006, Neuron.

[16]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[17]  Ronald L. Davis,et al.  Drosophila α/β Mushroom Body Neurons Form a Branch-Specific, Long-Term Cellular Memory Trace after Spaced Olfactory Conditioning , 2006, Neuron.

[18]  L H Schaefer,et al.  Structured illumination microscopy: artefact analysis and reduction utilizing a parameter optimization approach , 2004, Journal of microscopy.

[19]  Jerome Mertz,et al.  Optically sectioned fluorescence endomicroscopy with hybrid-illumination imaging through a flexible fiber bundle. , 2009, Journal of biomedical optics.

[20]  M. Heisenberg.  Mushroom body memoir: from maps to models , 2003, Nature Reviews Neuroscience.

[21]  T. Wilson,et al.  Method of obtaining optical sectioning by using structured light in a conventional microscope. , 1997, Optics letters.

[22]  A Borst,et al.  Drosophila mushroom body mutants are deficient in olfactory learning. , 1985, Journal of neurogenetics.

[23]  A. Nakano.  Spinning-disk confocal microscopy -- a cutting-edge tool for imaging of membrane traffic. , 2002, Cell structure and function.

[24]  T Nielsen,et al.  High efficiency beam splitter for multifocal multiphoton microscopy , 2001, Journal of microscopy.

[25]  Gilles Laurent,et al.  Transformation of Olfactory Representations in the Drosophila Antennal Lobe , 2004, Science.

[26]  Karel Svoboda,et al.  Stereotyped Odor-Evoked Activity in the Mushroom Body of Drosophila Revealed by Green Fluorescent Protein-Based Ca2+ Imaging , 2004, The Journal of Neuroscience.

[27]  G. Rubin,et al.  Refinement of Tools for Targeted Gene Expression in Drosophila , 2010, Genetics.

[28]  J. B. Duffy,et al.  GAL4 system in drosophila: A fly geneticist's swiss army knife , 2002, Genesis.

[29]  Sreekanth H. Chalasani,et al.  Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators , 2009, Nature Methods.

[30]  F. Del Bene,et al.  Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy , 2004, Science.

[31]  T. Ebner,et al.  Use of voltage-sensitive dyes and optical recordings in the central nervous system , 1995, Progress in Neurobiology.

[32]  Jerome Mertz,et al.  Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection. , 2010, Journal of biomedical optics.

[33]  Ryohei Kanzaki,et al.  Reconstructing the Population Activity of Olfactory Output Neurons that Innervate Identifiable Processing Units , 2008, Frontiers in neural circuits.

[34]  G. S. Kino,et al.  A Real-Time Confocal Scanning Optical Microscope , 1987, Other Conferences.

[35]  P. Stokseth.  Properties of a Defocused Optical System , 1969 .

[36]  Brendon O. Watson,et al.  SLM Microscopy: Scanless Two-Photon Imaging and Photostimulation with Spatial Light Modulators , 2008, Frontiers in neural circuits.