<ul><li><p>November/December 2011 UK Issue 149</p><p>Plasma FIB Analysisof ThroughSilicon Vias p9</p><p>Secondary ElectronAtomic Imagingin S/TEM pS5</p><p>Variable PressureSEM ofPlant Stigmata p21</p><p>High ThroughputAutomatedS/TEM pS11</p></li><li><p>Ensure your slides are never again lost, damaged or forgotten. </p><p>With the Olympus VS120 slide scanning and archiving system, you can turn your entire slide collection into a digital data mine, creating high resolution images of each slide that can be browsed virtually just as if you were sat at the microscope. You can share information with colleagues across the world or in the next room, at any time. And with the new fluorescence module you can permanently capture your data before your work can fade away. </p><p>For further information please visit: www.microscopy.olympus.eumicroscopy@olympus-europa.com</p><p>Keeping your slides, exactly as you know them.The Olympus VS120 virtual slide system. Digital slides at the highest resolution.</p><p>CIRCLE NO. 2 OR ONLINE: www.microscopy-analysis.com</p></li><li><p>REGULARS4 Dates for the Diary</p><p>23 People &amp; Places</p><p>26 Literature Highlights</p><p>29 Whats New</p><p>41 Reader Enquiry form</p><p>MICROSCOPY AND ANALYSIS NOVEMBER 2011 3</p><p>CONTENTS</p><p>www.microscopy-analysis.com</p><p>See the Microscopy and Analysis online Diary for a full listing. Entries for the January issue are due 1 December. Email to: editor@microscopy-analysis.com </p><p>dates for the diaryDIARY</p><p>MICROSCOPY AND ANALYSIS NOVEMBER 20114</p><p>NOVEMBER 20118-11 Gatan EELS School, Austrian Centre for Electron Microscopy and </p><p>Nanoanalysis, TU Graz, Austriawww.felmi-zfe.tugraz.at www.gatan.com/company/news/news08291102.php</p><p>10 Electron Microscopy in Materials Science, Eindhoven University of Technology, The Netherlandshttp://nvvmmaterials2011.chem.tue.nl/ </p><p>12-16 Neuroscience 2011: Washington DC, USAwww.sfn.org</p><p>14-18 NvVM European School on Nanobeams , Centre de Recherche Public - Gabriel Lippmann, Belvaux, Luxembourg www.nanobeams.org </p><p>16 Cryo Microscopy Group Annual Meeting, Boots Science Building, School of Pharmacy, University Park, Nottingham University, UKwww.cryomicroscopygroup.org.uk/CMG2011.html</p><p>22-23 Bruker Scanning Probe Microscopy Conference and Users Meeting, University of Manchester, UKwww.bruker-axs.com drew.murray@bruker-nano.com</p><p>28-2/12 2011 MRS Fall Meeting, Boston, Massachusetts, USAwww.mrs.org</p><p>DECEMBER 20113-4 Functional Optical Imaging, Univeristy of Nottingham Ningbo Campus,</p><p>Chinawww.nottingham.ac.uk/ibios/index.php?page=foi-11</p><p>3-7 American Society for Cell Biology 51st Annual Meeting, Denver, Colorado, USAwww.ascb.org</p><p>JANUARY 20124-7 Winter School on High-Resolution Electron Microscopy, Arizona State </p><p>University, Tempe, AZ, USAwintershcool@asu.edu http://le-csss.asu.edu/winterschool </p><p>FEBRUARY 20125-9 APMC 10: 10th Asia-Pacific Microscopy Conference; </p><p>ICONN 2012: International Conference on Nanoscience and Nanotechnology; </p><p>ACMM 22: Australian Conference on Microscopy and Microanalysis, Perth, Australiawww.apmc-10.org www.iconn-2012.org www.acmm-22.org</p><p>25-29 56th Biophysical Society Annual Meeting, San Diego, California, USAwww.biophysics.org/2012meeting/Main/tabid/2386/Default.aspx </p><p>MARCH 201211-16 Pittcon 2012, Orlando, FL, USA</p><p>www.pittcon.org18-23 Course in Cryotechniques for Electron Microscopy, Rothamsted </p><p>Research, Harpenden, UKwww.rms.org.uk/events/Forthcoming_Events/CoolRunnings</p><p>APRIL 20121-4 Focus on Microscopy 2012; 25th International Conference on 3D Image </p><p>Processing in Microscopy; 24th International Conference on Confocal Microscopy, Singaporewww.focusonmicroscopy.org</p><p>9-13 MRS 2012 Spring Meeting and Exhibition, San Francisco, CA, USAwww.mrs.org/spring2012</p><p>17-20 Analytica 2012: 23rd International Trade Fair for Laboratory Technology, Analysis and Biotechnology and analytica Conference, Munich Trade Fair Centre, Munich, Germanywww.analytica.de</p><p>MAY 20129-11 AMTC3: 3rd International Symposium on Advanced Microscopy and </p><p>Theoretical Calculations, Nagaragawa Convention Center, Japanwww.congre.co.jp/amtc3/</p><p>21-25 IFES2012: 53rd International Field Emission Symposium, University of Alabama, Tuscaloosa, AL, USAwww.ifes2012.au.edu</p><p>24-25 SCUR 2012: 39th Annual Meeting of Society for Cutaneous Ultrastructure Research, Lyon, Francehttp://orgs.dermis.net/content/e04scur/e03meetings/e770/e1074/index_ger.html </p><p>JUNE 20123-15 Microscopy School, Lehigh University, Bethlehem, PA, USA</p><p>Sharon Coe: sharon.coe@lehigh.edu www.lehigh.edu/microscopy</p><p>3 Introduction to SEM and EDS for the New Operator4-8 SEM and X-Ray Microanalysis11-14 Focused Ion Beam: Instrumentation and Applications11-15 Problem Solving with SEM, X-Ray Microanalysis and Electron Backscatter Patterns11-15 Quantitative X-Ray Microanalysis: Problem Solving with EDS and WDS Techniques11-15 Scanning Transmission Electron Microscopy: From Fundamentals to Advanced Applications</p><p>25-27 Abercrombie Cell Biology Symposium, Oxford, UKBSCB: www.bscb.org</p><p>27-30 CARS 2012: Computer Assisted Radiology and Surgery, 26th Intl Congress and Exhibition, Joint Congress of CAR / ISCAS / CAD / CMI / EuroPACS, Congress Palace, Pisa, Italywww.cars-int.org</p><p>JULY 20122-6 Electron Microscopy Summer School, University of Leeds, UK</p><p>RMS: www.rms.org/events 2-6 Light Microscopy Summer School, University of York, UK</p><p>RMS: www.rms.org/events 4-6 Optics Within Life Sciences (OWLS), Genoa, Italy</p><p>Organised by Italian Institute of Technology www.owls2012.org9-13 Inter/Micro: 63rd Annual Applied Microscopy Conference, Chicago, IL, </p><p>USAwww.mcri.org</p><p>11-13 CryoElectron Microscopy Short Course, University of Minnesota, MN, USANanostructural Materials and Processes Program of IPrime: mccormic@umn.edu www.iprime.umn.edu </p><p>29-2/8 Microscopy &amp; Microanalysis 2012, Phoenix, Arizona, USAwww.microscopy.org</p><p>AUGUST 20126-10 ULTRAPATH XVI: Conference on Diagnostic Electron Microscopy, Basic </p><p>Research and Oncology, Regensburg, Germanywww.ultrapath.org</p><p>26-30 14th International Congress of Histochemistry and Cytochemistry, Kyoto, Japan www.acplan.jp/ichc2012/</p><p>SEPTEMBER 2012 16-21 European Microscopy Congress, Central Convention Complex, </p><p>Manchester, UKwww.rms.org.uk</p><p>OCTOBER 201213-17 Neuroscience 2012, New Orleans, Louisiana, USA</p><p>www.sfn.org </p><p>NOVEMBER 201226-30 2012 MRS Fall Meeting &amp; Exhibition, Boston, Massachusetts, USA</p><p>www.mrs.org/fall2012</p><p>DECEMBER 201215-19 ASCB 52nd Annual Meeting, San Francisco, CA, USA</p><p>www.ascb.org</p><p>MARCH 201317-21 Pittcon 2013, Philadelphia, PA, USA</p><p>www.pittcon.org</p><p>APRIL 20131-5 2013 MRS Spring Meeting &amp; Exhibition, San Francisco, CA, USA</p><p>www.mrs.org/spring2013</p><p>AUGUST 20134-8 Microscopy &amp; Microanalysis 2013, Indianapolis, IN, USA</p><p>www.microscopy.org</p><p>NOVEMBER 20139-13 Neuroscience, San Diego, California, USA</p><p>www.sfn.org</p><p>LITERATURE HIGHL IGHTS</p><p>literature highlights' '! '</p><p>A large array of sub-10-nm sin-gle-grain gold nanodots for usein nanotechnology is describedby Nicolas Clment and col-leagues at the Institut dElec-tronique Microlectronique etNanotechnologie, CNRS, Univer-sity of Lille, France [Small7(18):2607-2613, 2011].A uniform array of single-</p><p>grain gold nanodots, as small as5-8 nm, was be formed on siliconusing e-beam lithography. Theas-fabricated nanodots wereamorphous, and thermal annealing converted themto pure Au single crystals covered with a thin SiO2layer. These findings were based on physical mea-surements by AFM, atomic-resolution STEM, andchemical techniques using energy dispersive X-rayspectroscopy. The authors demonstrated the forma-tion by e-beam lithography of sub-10-nm Au dotswith small dispersion and perfect alignment. Suchprecise formation of small dots enabled them to iden-</p><p>(a) STEM image showing the bulk silicon (Si), five annealed dots (Au), carbon layer (C), and platinum layers. (c) Coloured STEM image of a single annealed nanodot (260C, 2 h). Reproduced with permission, Copyright 2011 Wiley-VCH Verlag GmbH &amp; Co. KGaA, Weinheim. </p><p>tify the critical size that determines whether a dot iscomposed of single or multiple crystal domains.Moreover, they showed that annealing at moderatetemperature can convert Au dots from amorphous tosingle-crystalline, and then they were covered with athin SiO2 layer. After easy removal of the SiO2 (diluteHF etching), these nanodots can be used as electrodesfor the characterization of organic self-assembledmonolayers (SAMs) with less than 200 molecules.</p><p>' ! 'A miniature stage device to overcome resolutionanisotropy in fluorescence light microscopy isdescribed by Florian Staier and colleagues at theKirchhoff Institute for Physics, University of Heidel-berg, Germany [Rev. Sci. Instrum. 82:093701, 2011].To overcome the limitation of fluorescence micro-</p><p>scopes in anisotropic optical resolution or point local-ization precision micro-axial tomography was usedwhich allowed object tilting on the microscope stageand led to an improvement in localization precisionand spatial resolution. A glass fiber was placed in theobject space of the microscope lens and its rotationcontrolled by a miniaturized stepping motor. By Testparticles were fixed onto the glass fiber, opticallylocalized with high precision, and automaticallyrotated to obtain views from different perspectiveangles from which distances of corresponding pairs ofobjects were determined. From these angle depen-dent distance values, the real 3D distance was calcu-lated with a precision in the ten nanometer range(corresponding here to an optical resolution of 10-30nm) using standard microscopical equipment. As aproof of concept, the spindle apparatus of a maturemouse oocyte was imaged during metaphase II mei-otic arrest under different perspectives.</p><p>%# $!</p><p>A technique for the use of two fluorophores in stim-ulated emission depletion (STED) microscopy of livingcells is reported by Patrina Pellett and co-workers atthe Department of Cell Biology, Yale School of Medi-cine, CT [Biomedical Optics Express 2(8):2364-2371,2011]. Current applications of STED microscopy havebeen limited to single colour imaging of living cellsand multicolour imaging in fixed cells. However, tostudy active processes, such as protein interactions, atwo-colour STED imaging technique is needed in liv-ing cells. This was achieved for the first time by theauthors: the key to their success was in overcomingthe challenges in labeling target proteins in livingcells with dyes optimal for two-colour STEDmicroscopy. By incorporating fusion proteins, theresearchers were able to improve the targetingbetween the protein and the dye, effectively bridgingthe gap. This allowed the researchers to achieve reso-lutions of 78 nm and 82 nm for 22 sequential two-colour scans of epidermal growth factor and its recep-tor in living cells. </p><p>' #'! 'The three-dimensional point spread function of anaberration-corrected scanning transmission electronmicroscopy (STEM) has been simulated and experi-mentally tested by Andrew Lupini and Niels de Jongeat the Oak Ridge National Laboratory, TN [Microscopyand Microanalysis 17:817-826, 2011].Aberration correction reduces the depth of field in</p><p>STEM and thus allows three-dimensional imaging bydepth sectioning. This imaging mode offers thepotential for sub-ngstrom lateral resolution andnanometer-scale depth sensitivity. For biological sam-ples, which may be many m across and where highlateral resolution may not always be needed, opti-mizing the depth resolution even at the expense oflateral resolution may be desired, aiming to imagethrough thick specimens. Although there has beenextensive work examining and optimizing the probeformation in two dimensions, there is less knownabout the probe shape along the optical axis. The authors examined the probe shape in 3D in an</p><p>attempt to better understand the depth resolution in</p><p>'!!''!A fractal dimension analysis and mathematical mor-phology of structural changes in actin filamentsimaged by electron microscopy is reported by Yoshi-taka Kimori et al at the National Institutes of NaturalSciences, in Tokyo [Journal of Structural Biology176(1):1-8, Oct 2011].The authors examined structural changes of actin</p><p>filaments interacting with myosin visualized by quickfreeze deep-etch replica EM by using a new methodof image processing and analysis based on mathe-matical morphology. To quantify the degree of struc-tural changes, two characteristic patterns wereextracted from the EM images: the winding patternof the filament shape (WP) reflecting flexibility of the</p><p>' !#'#!A. Beltrn and colleagues at the Department of Mate-rials Sciences, University of Cadiz, Spain, report thatthree dimensional atom tomography resolves thequantum ring morphology of self-assembled GaSbburied nanostructures [Ultramicroscopy 111(8):1073-1076, July 2011].Unambiguous evidence of ring-shaped self-assem-</p><p>bled GaSb nanostructures grown by molecular beamepitaxy is presented on the basis of atom-probetomography reconstructions and darkfield transmis-sion electron microscopy imaging. From atom-probetomography compositional distribution has beenobtained. The GaAs capping process causes a strongsegregation of Sb out of the center of GaSb quantumdots, leading to the self-assembled GaAsxSb1-x quan-tum rings of 20-30 nm in diameter with x~0.33.</p><p>filament, and the surface pattern of the filament (SP)reflecting intramolecular domain mobility of the actinmonomers in the filament. EM images were processedby morphological filtering followed by box-countingto calculate the fractal dimensions for WP (DWP) andSP (DSP). The result indicated that DWP was largerthan DSP irrespective of the state of the filament(myosin-free or bound) and that both parameters formyosin-bound filaments were significantly largerthan those for myosin-free filaments. This work is thefirst quantitative insight into how conformational dis-order of actin monomers is correlated with themyosin-induced increase in flexibility of actin fila-ments along their length.</p><p>this mode. They present examples of how aberrationschange the probe shape in three dimensions, and itwas found that off-axial aberrations may need to beconsidered for focal series of large areas. It was shownthat oversized or annular apertures theoreticallyimprove the vertical resolution for three-dimensionalimaging of nanoparticles. When imaging nanoparti-cles of several nanometers in size, regular scanningtransmission electron microscopy can thereby be opti-mized such that the vertical full-width at half-maxi-mum approaches that of the aberration-correctedSTEM with a standard aperture.</p><p>MICROSCOPY AND ANALYSIS NOVEMBER 201126</p><p>CIRC</p><p>LENO. 1 O</p><p>RONLINEwww.microscop</p><p>y-analysis.com</p><p>The cover shows a confocal image (top) of aDrosophila embryo at stage 11, expressing the trachealmarker trh-LacZ (Cy3, red) and the cell membranemarker Dlg (Alexa 488, green). The enlarged view(below) shows invaginating tracheal placodes in X-Y(left) and Y-Z (right) projections. (Courtesy of DrTakefumi Kondo and Dr Shigeo Hayashi at theLaboratory for Morphogenetic Signalling, RIKEN Centrefor Developmental Biology, Japan.)</p><p>The new UPLSAPO30xS and UPLSAPO60xS siliconoil-immersion objectives from Olympus are ideal for livecell experiment..</p></li></ul>