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EUROPEAN RESEARCH EDITION

09 NANOTUBES AND SYNAPSESIN IMMUNE CELL COMMUNICATION

Confocal Laser Scanning Microscopy

12 SEEING LIFE BEGINDynamic Imaging Sheds Light onEmbryonic Development Processes

14 THE STEREOMICROSCOPE AS A 3D MEASUREMENT DEVICE

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reSolution Newsletter2_01_05 14.07.2005 9:11 Uhr Seite 1

Dear ReaderMeeting the current and future needs of Leicacustomers was the driver for the reorganisationof Leica’s European Sales Divisions last year. InLife Science Research, Leica considered thechanges to be a pre-requisite for commercial

success as this could only be achieved by clearly understanding theneeds of our customers and by providing leading edge products tosatisfy those needs.

Therefore, in July 2004, Leica formed a new European ResearchSales Division totally dedicated to addressing Life Science Researchin the area of Microscopy and High End Imaging Systems. The timingof the reorganisation was also influenced by the simultaneous intro-duction of a new generation of Microscopy Systems offering previ-ously unavailable benefits to key areas of life sciences especially, butnot only, in the area of Live Cell Imaging and Stem Cell Research.

The development and introduction of this new generation of prod-ucts, from Confocals to quantitative Widefield and advanced Fluores-cence Systems, is ongoing with more ground breaking and uniqueproducts coming to market in the near future.

In addition to product offerings, Leica also has a wealth of applica-tions knowledge and high end imaging expertise, all of which may beof value to European scientists.

As a vehicle to bring news of new product developments to the scien-tific community and to be able to share our knowledge and expertisewe have decided to produce a European edition of our current Reso-lution magazine specifically for Life Science Research.

This magazine will develop as we respond to reader feedback andwill be published regularly from now on. I do hope you enjoy our firstedition and it brings you some real value.

John BuckleyDirector European Research Sales DivisionLeica Microsystems

E D I T O R I A L

Leica AF6000

Leica Microsystems introduces the new Leica AF6000system for fluorescence imaging and live cell analy-sis. This extremely flexible system can be configuredto all requirements for use in highly analytical appli-cations, turning brilliant fluorescence images intosignificant data.

Please see page 8 for further details.

Leica TCS SP2 SESensitivity Enhanced for Brilliant Images

Sensitivity is the basis for high imaging quality. The new Leica spectralconfocal TCS SP2 SE uses technologies to further improve image qualityand significance of data. In addition to the well proven filter free AOBS®

beam splitting and SP® tunable detection, the SE version is equippedwith extra sensitive PMTs, and allows optional APD imaging at very high quantum efficiency and very low dark noise for brilliantimages. The Super VISIR coating drastically improves transmission,especially in the infrared range for MP (multiphoton microscopy). The TCS SP2 SE offers better light sources as well: double power 405nm laser for DAPI, Hoechst, paGFP; and a high energy Ar-laser for FRAP experiments.

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ORIA

LreSolution Newsletter2_01_05 14.07.2005 9:11 Uhr Seite 2

NEW PRODUCTS

Leica DMI3000 B and DMI4000 B complementthe Leica DMI6000 BThe correct instrument for any biomedical application

New micromanipulation systemby Leica and Eppendorf

New system for fluorescence imagingand live cell analysis: AF6000

APPLICATION REPORTS

Confocal Laser Scanning Microscopy Nanotubes and Synapses in Immune CellCommunication

Dynamic imaging sheds lighton embryonic development processes:Seeing Life Begin

The stereomicroscope as a 3D measurement device

The world’s onlyinteractive Leica 3D system for microscopy

Leica MacroFluo™ – the new precisionin fluorescence macroscopy

EUROPEAN RESEARCH EVENTS

CONTEST:Your opinion is valuable!

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Welcome to the first publication of reSOLUTION – European Research Edition!

This magazine is for you, dear customer. We have created this forum to provide you withproduct information and many of the upcoming European Leica Microsystems events overthe next few months. Our goal is to publish this newsletter 2-3 times a year.Also in this first edition, we will present to you two researchers using Leica Microsystemsproducts in their everyday work: First, Dr. Daniel Davis (Imperial College, London) who haskindly given us a recent article on Molecular Immunology. Secondly, you can read aboutdevelopmental biology on zebrafish from the lab of Dr. Reinhard Koester (GSF Neuherberg,Germany).Finally, a contest where you can win great prizes, just for giving us your opinion, awaits youalong with the calendar of events. So enjoy the newsletter, and I look forward to hearingfrom you.

Paul Wismer, European Marketing Manager, Research, Leica Microsystems


Leica Microsystems has introduced a new line ofinverted research microscopes for the discerninglive cell researcher: The Leica DMI Series. Brilliance,convenience, integration, automation, and flexibilitycharacterize the innovations.

“Simply Microscopy“ is the central theme for Leica’sDMI microscopes. This phrase describes intelligentautomation of the microscope, with the result that aresearcher can concentrate fully on his or her experi-ment and not on microscope functions.

The Leica DMI research series, with its DMI3000 B,DMI4000 B, and DMI6000 B models, offers the rightsolution for all live cell applications. The DMI familyincludes fully automated versions, as well as ’coded’systems that make it easier to select the correct com-ponents, and document your settings. A completely

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N E W P R O D U C T S

Range Expansion of Inverted Research Microscopes

Leica DMI3000 B and DMI4000 B Complementthe Leica DMI6000 B

manual stand is also available for cost-conscioususers. The Leica DMI3000 B is designed for cus-tomers that work exclusively with transmitted lightmethods. The new Leica DMI3000 B and DMI4000 Binverted microscopes not only exceed the latest tech-nical standards, they also fulfil all ergonomic require-ments to provide comfortable use during long periodsof time at the microscope. In addition to live cell re-search, Leica DMI microscopes are ideal for routineexaminations such as scanning cell or tissue cul-tures.

Highly complex microscope processes are not onlymotorised in the DMI6000 B and DMI4000 B, but fullyautomated, so that users can concentrate fully ontheir research work. Leica’s intelligent automationstill leaves room for individual requirements, how-ever: manual intervention is possible at any time.


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Through improved optics and reduced back-ground radiation, the Leica DMI4000 B andDMI6000 B achieve especially brilliant fluo-rescence. The fluorescence turret isequipped with up to six filter cubes thatcan be moved into position quickly,without vibration, at the touch of abutton.

The fluorescence axis also featuresan ultrafast, internal filter wheel (IFW)that supports the excitation of fluo-rochromes in less than 20 milliseconds.The Fluorescence Intensity Manager(FIM) reduces light stress for living cellsand the bleaching of fluorochromes – the in-tensity of the excitation light can be reduced in areproducible manner while the quality of the fluores-cence is improved.

The integrated, motorised Excitation Manager (Ex-Man) balances a variety of emission intensities tosubsequently compensate for specimen artifacts.This innovative combination is truly unique. The microscopes are also designed to be completelylight-tight, making it possible to mount even highlysensitive cameras on one of the four ports, withoutadversely affecting results. A versatile line of acces-sories, tailored to the microscope, create ideal condi-tions for live cell observation. These include incuba-tors and heating/cooling devices, to provide idealphysiological conditions for cells at all times in regardto temperature, carbon dioxide, and oxygen ratios.Particularly for micromanipulation, Leica has designedespecially narrow specimen stages that allow manipu-lators and other peripheral devices to be positionedclose to the specimen. > VN




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Range Expansion of Inverted Research Microscopes

The Correct Instrument forAny Biomedical Application

Leica Microsystems Inc, the expert in innovative,high-tech, precision optical instrument design, hasexpanded its line of DMI inverted research micro-scopes with the new Leica DMI3000 B and LeicaDMI4000 B models. “Simply Microscopy” is the centraltheme for Leica’s DMI inverted microscope series,that focuses on simple operation through intelligentautomation.

The Leica DMI inverted research series, with itsDMI3000 B, DMI4000 B and DMI6000 B models, offersa suitable solution for all live cell applications. Highlycomplex processes are not only motorised in theDMI6000 B and DMI4000 B, but fully automated, al-lowing users to concentrate completely on their re-search work. Leica’s intelligent automation stillleaves room for individual requirements, however,and manual intervention is possible at any time.

The Leica DMI family includes fully automated ver-sions as well as ’coded’ systems that make it easier toselect the correct components, and document yoursettings. A completely manual stand is also availablefor cost-conscious users. The Leica DMI3000 B is de-signed for customers that work exclusively withtransmitted light methods.

The new Leica DMI3000 B and DMI4000 B invertedmicroscopes not only exceed the latest technicalstandards, they also fulfil all ergonomic require-ments to provide comfortable use during long peri-ods of time at the microscope. The DMI series isdesigned for the study of live cells, micromanipula-tion, electrophysiology, and is also suitable for rou-tine examinations such as scanning cell or tissuecultures. > VN



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New Micromanipulation System

New System by Leica and EppendorfLeica Microsystems Inc. and Eppendorf present thesecond generation of their micromanipulation line. Asthe successor to the Leica AS TP Transgenic Platform,the Leica AM6000 is based on state-of-the-art technol-ogy – the Leica DMI6000 B inverted digital microscope.AM6000 users will benefit from all of the advantages ofthis fully automated research instrument.

The comprehensive integration of microscope and mi-cromanipulators in the new manipulation system offersa wide range of new possibilities."Simply Microscopy"is the central theme for the Leica DM line of researchmicroscopes that focuses on simple operation throughintelligent automation.

The complete Leica AM6000 system is controlledfrom multifunction panels that cover not only micro-

manipulation, but also the most important microscopefunctions. This integrated system accelerates workprocesses and enhances efficiency.

In addition to all of the customary functions of NK2-generation Eppendorf manipulators, the panels alsofeature focus handwheels and function buttons forbrightness, objective selection, and the magnificationchanger. Objectives can be associated with contrastmethods, stored with the magnification changer, andrecalled as required.

The Leica AM6000 is designed especially for trans-genics and ICSI applications. > VN



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From overlaying multi-channel images to z stackingand time lapse features, a wealth of solutions are in-cluded as standard for image documentation, quan-tification enhancement and analysis. Designed tocompletely harmonise microscope, camera and ap-plication, the Leica AF6000 is compatible with a widerange of Leica Microsystems fluorescence researchmicroscopes and is well matched to both upright andinverted instruments.

The AF6000 is based on a new software platformwhich is shared by all imaging products of Leica Micro-systems. The simplicity of its highly intuitive interfacewith guided workflow ensures fast and effortlesshandling and guarantees brilliant results in a shorttime.

New AF6000 System

New System for FluorescenceImaging and Live CellAnalysis


A P P L I C AT I O N R E P O R T

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Many of the key cell surface molecules involved inimmune cell surveillance are now identified and oneimportant new scientific frontier is to understand whereand when each protein-protein interaction occurs toregulate cell functions. It is well established, for ex-ample, that white blood cells called Natural Killer(NK) cells recognize and kill tumor or virus-infectedcells via different activating receptors or by detectinga loss of self-protein using inhibitory receptors buthow cell-cell interactions over a few minutes leads tothe decision whether or not a cell is diseased is farfrom understood. Thus, imaging has a major role toplay in contemporary cell biology and one interestingtheme to emerge is that immune cell communicationis often accompanied by the segregation of proteinsinto micrometer-scale domains at an intercellularcontact or immunological synapse.

Patterns of proteins that form at immune synapsesresult from intimate cooperation between severalfactors: Inter- and intramembrane protein binding,membrane mechanics, and cytoskeletal rearrange-ments all lead to a lateral sorting of proteins withintheir respective fluid membranes. Immune synapsesextend several micrometers across intercellular con-tacts and, at least in some circumstances, specificpatterns of proteins correlate with the ensuing intra-cellular signalling and cellular effector functions.

Recently, for example, we have examined the mem-brane localisation of proteins that are up-regulatedupon infection or tumour transformation and can acti-vate human NK cells [Eleme et al., J. Exp. Med., 2004,199:1005]. By using laser scanning confocal micro-scopy and electron microscopy we showed thatthese stress inducible ligands, MICA and ULBP1, 2, 3,are expressed in specific membrane microdomains(lipid rafts). The molecular mechanism by whichthese proteins are organised at the cell surface in-volves covalent attachment to specific lipids. It isknown that viruses can prevent cell surface expres-sion of the NK cell activating proteins to evade NKcell activation. Perhaps also some viruses or tumoursevolved mechanisms to destabilize NK activation by

Dan Davis obtained a BSc. in Physics from the Uni-versity of Manchester in 1992 and a PhD. in Physicsin 1995, working with David Birch at StrathclydeUniversity. Then, as an Irvington Institute ResearchFellow, he studied immunology with Jack Stro-minger at Harvard University. In December 1999 hereturned to London to establish his own researchgroup at Imperial College. In 2000, he won theOxford University Press/Times Higher EducationSupplement Science Writing Prize and, in 2002,was awarded a European Molecular Biology Organ-isation Young Investigator award. Dan is currentlyauthor or co-author of about 60 scientific papers inphotophysics and immunology, including the firstdescription of a Natural Killer cell immune synapse,studies of proteins transferring between immunecells, and observations of membrane nanotubesconnecting immune cells.

Dr. Daniel M. DavisMolecular ImmunologyImperial College LondonDepartment of Biological Sciences

Confocal Laser Scanning Microscopy

Nanotubes and Synapses inImmune Cell Communication



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disrupting the cell surface organisation of these NKcell ligands. Our images also revealed an unexpectedintercellular transfer of proteins during immune sur-veillance [Vanherberghen et al., Proc. Natl. Acad.Sci., 2004, 101:16873].

The functional consequences of the intercellulartransfer of proteins during immune surveillance arelargely unknown but perhaps, for example, tagginghealthy cells could eliminate repeated scanning ofthe same cell.

In another recent article [Önfelt et al., J Immunol,2004, 173:1511], we built upon work published in anearlier neuroscience paper [Rustom et al., Science,2004, 303:1007], by presenting evidence of a novelmechanism for intracellular communication in immun-ology. Nanotubular highways, or membrane nano-tubes, were seen to connect multiple cells togetherand were readily formed between varieties of im-mune cell types. Nanotubes could be created upondisassembly of immune synapses, as cells moveapart, and so nanotube networks could be assembledfrom transient synapses. It seems that nanotubes cantransport membrane material along the surface of thetube as well as inside the tube. Therefore, nanotubesmay provide a mechanism for the transport of cell

surface proteins following the formation of an im-mune synapse. We were able to make this discoveryby using single photon-excitation resonance scan-ning confocal microscopy to image immune cell inter-actions in vitro very rapidly.

The immune synapse is likely to have multiple func-tions with varying degrees of importance for differentcell-cell interactions. In addition to controlling intra-cellular signals, the structure of the immune synapsemay also play a role in directing the secretion of cyto-kines or lytic granules towards the target cell. To un-derstand immune surveillance further, it will help toimage several protein species simultaneously, withsingle-molecule resolution, second-scale time resol-ution, and with objective automated analysis of pro-tein distribution. We also need to make real-timeobservations of synapse formation in living tissues,perhaps for example, using two-photon confocal mi-croscopy. Our understanding of the spatio-temporalaspects of immune cell communication is only justbeginning and the application of novel imaging tech-nologies to this area of research is more than likely tothrow up further unexpected observations in the nearfuture.

The NK cell immunological synapse. Figure shows a B-cell line (221/Cw6-GFP) transfected to express GFP-tagged class I MHC protein (green) surrounded by three NK cells (YTS/KIR2DL1)transfected to express an inhibitory receptor. Lysotracker staining acidic organelles is shown in red and DNA, stained by Hoechst, is shown in blue. Clusters of GFP-tagged class I MHC proteinat the intercellular contacts mark the immune synapses formed.



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Bebhinn Treanor and Bjorn Önfelt use a Leica resonance-scanning confocal microscope to examine immune synapse assembly. Photo by David Bacon.

This article was published by:

Gemma L. Buckland, Bjorn Önfelt & Daniel M. Davis

Division of Molecular and Cellular Biology, Sir Alexander Fleming Building,Imperial College London, UK. E-mail: [email protected]

Further Reading

DM Davis, Trends in Immunology 2002, 23: 356DM Davis & ML Dustin, Trends in Immunology 2004,25: 323



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Without dynamic imaging methods, capturing embryonic develop-ment processes would be unthinkable. To automate laborious time-lapse experiments on living zebrafish embryos, the developmentalbiologists at the GSF (National Research Center for Environment andHealth) in Neuherberg, Germany, tested a complete software-con-trolled system developed by Leica Microsystems. The motorised fluo-rescence stereomicroscope, fluorescence imaging software and digitalcamera work together in precision, easily handling even complextime-lapse and Z-stack experiments with quick filter change routinesand deconvolution. Comprehensive all-around testing as well as spe-cific tests to determine the precision of the interaction between hard-ware and software made it obvious that, in every respect, the LeicaSystem enriches everyday scientific work in a zebrafish neuroimaginglaboratory. It is a cost-efficient alternative to expensive instrumentsfor intravital microscopic imaging.

Fig. 1: Time-lapse images of a zebrafish embryo with injected contrast medium

The zebrafish – transparency in the world of biology

Martin Distel of the Zebrafish Neuroimaging GroupReinhard Köster at the GSF Neuherberg uses the ze-brafish as a model organism for intravital studies ofgenetic development. Native to India's Ganges river,this popular research subject is highly sought after inbiology labs around the world. A look through the mi-croscope reveals the secret of its popularity: the ze-brafish embryo is transparent, making it possible toobserve firsthand the development of organs, such asthe eye, as it grows.

Leica’s complete system – put to the test

To efficiently capture images of development pro-cesses, the software-controlled system from LeicaMicrosystems was tested in various experiments onliving zebrafish embryos. The FW4000 software wasable to reliably automate all time-lapse experimentsin bright-field and fluorescence modes and couldhandle the imaging of rapidly occurring processes,such as the various phases of the heartbeat (Fig. 1).

Even complex time-lapse experiments with threefluorescence filters functioned perfectly (Fig. 2). UV-convertible pigments in cells of a zebrafish embryowere excited in succession with GFP3, DAPI andTexas Red. The motorised filter change was fast andso free of vibration that high-quality images could beobtained without any pixel shift.

The supreme discipline: complex experiments with filter change, Z-stack and deconvolution

The motorised focusing drive on the Leica MZ16 FAallows automated imaging in various planes along theZ-axis. In the test, a Z-stack in 15 planes was taken of

Dynamic Imaging Sheds Light on Embryonic Development Processes

Seeing Life Begin



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a doubly marked zebrafish embryo (Fig. 3). With theFW4000 software, the focal displacements and thechange routine for the two fluorescence filters func-tioned reliably and without vibration. Then, the im-ages from the individual channels (GFP3 and TexasRed) for each Z-plane were composed into a singleimage using the FW4000 software (Fig. 3c). This canproduce a blurry image, as is the case here, becauseregular images taken with a stereomicroscope con-tain a lot of information from areas outside of thefocal plane. Also, the overlapping “red and green” in-formation creates a yellow colour that misleadinglyindicates a colocalisation of nuclei and cellular mem-branes. The Leica Deblur imaging software providesthe solution to this problem with its deconvolutionfunction, which assigns non-focused light signals totheir original focal plane. The deconvolution workedvery well. Not only was the composite sharper (Fig.3d), the subcellular structures also appeared in con-vincingly high resolution, and the yellow areas dis-appeared almost completely.

Experiments were convincing

The software package from Leica Microsystems isvery promising indeed for automating time-lapse and Z-stack experiments, and in combination with the

MZ16 FA fluorescence stereomicroscope, can open up new paths in develop-

ment research. In spite of its complexity, the software is quite easy to learn and use.

The quality of the images achieved and the perform-

ance of the completeLeica system convincedthe researchers at GSF

Neuherberg.> IH

Fig. 2: Complex time-lapse experiment with 3 fluorescence filters (GFP3, DAPI, Texas Red). Automated contrastmedium conversion. Embryo before, during and after irradiation

Fig. 3: Zebrafish embryo with multicolouredfluorescence-marked nuclei and cellularmembranes; level 8 of the Z-stacka) GFP3 channel with fluorescence-markedgreen cell nucleib) Texas Red channel with fluorescence-marked red cellular membranesc) Composite of a and b before deconvolu-tiond) After deconvolution. Yellow areas havedisappeared almost completely, i.e. the flu-orescence signals are correctly assignedto their focal plane.



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Introduction

Traditionally microscopists have used their stereomi-croscopes for stereoscopic viewing and 2D imaging.There are countless uses for the stereomicroscopeto examine the surfaces of specimen, but the chal-lenge has always been the microscope’s inability tomeasure (and even image) in 3D. In stereomi-croscopy the depth of focus is large, however no di-rect 3D visualisation and measurement are possiblethroughout this extended depth of focus. Conse-quently, when quantitative 3D surface analysis is de-sired, an alternative device is needed to supplementthe stereomicroscope, or other solutions have to beapplied. This simple limitation has posed an extremechallenge to scientists and their budgets of time,money…and results.

In this report a new 3D analysis capability in stereo-microscopy from Leica Microsystems is presented.Conventional stereomicroscopes can be trans-formed into true 3D measurement devices with fullanalytical capability to generate profile, roughness,area and even volumetric measurements. The Leica3D system is useful in all fields of science and tech-nology in which representation of spatial depth infor-mation means having more, and more valuable, infor-mation.

3D Stereo Reconstruction

Currently no real 3D analysis is commercially avail-able. This limitation is circumvented with the stand-alone software package Stereo Explorer from LeicaMicrosystems. This modular software automaticallycalculates a 3D data record from two partial imagesfor reconstruction, documentation, analysis andquantitative measurement of three-dimensional sur-faces. The realistic image, which appears in relief,makes it easier for the user to identify complex sur-faces, greatly improves education and training, andenables better diagnoses in technical, biological andforensic fields.

Stereopairs with excellent-quality, true-to-detail im-ages are an essential requirement for precise 3D re-

constructions. The integrated digital Leica IC 3Dstereo camera creates a perfect high-resolutionstereopair without the need for complex adjustments(Figure 1).

Conventional stereoscopic approaches require ac-curate optical setups in order to get acceptable re-sults. Developments by Leica Microsystems greatlyreduce the requirements of that because an auto-matic offset calculation and geometric correctionare performed. This calculation produces highly ac-curate and robust 3D surface reconstructions. Therelative height accuracy is better than 3% and a fullyautomatic calculation can be performed in less than100 seconds. The software can be applied to anystereomicroscope of the Leica Microsystems M-Series. The version for the Leica MZ16 A automatedstereomicroscope controls not only the Leica IC 3Dstereo camera, but also the motorised zoom and up-dates the data on the display. In addition, the autofo-cus module allows precision control of the motorisedfocus. These settings can be saved and retrieved asrequired. This makes work ergonomic and efficient,particularly during precise, repetitive experiments.

The Stereomicroscopeas a 3D Measurement Device

Fig. 1: The optical concept of 3D image capture: The Leica IC 3Ddigital stereo camera is integrated between the binocular tube andoptics carriers of a Leica M Series stereomicroscope and takes ahigh-resolution image of each optical beam path.



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Quantitative measurement

Once the surface information of the object is deter-mined, the 3D data can be visualised and analysed.All analysis routines are designed in a modular, intu-itive manner, and are supported with full windows re-porting conventions. The possible analytical capabil-ities comprise numerous calculations where the de-scription of such would go beyond the limits of thispublication. The profile analysis module permits theextraction of height profile along user defined pathsand can be used to calculate relative height mea-surements as shown in figure 2.

All established roughness, profile and waviness

measurements can be performed as well as variousstatistical calculations. The area analysis modulesimilarly calculates parameters such as the ratio oftrue-to-projected area, also termed the bearing areacurve. The volume analysis module as shown in fig-ure 3 enables direct volumetric measurements rela-tive to freely definable polygon lines.

Applications

The technology presented in this article can be ap-plied in any field of study where the surface structureof a specimen is important. This may include docu-mentation of the roughness of microscopic specimensfor affinity calculations, the analysis of fractured sur-faces from mechanical parts, or the volumes of im-

Fig. 2:Profile of a geological specimen

Fig. 3: Volume measurement in fractureanalysis. A 3D model visualises the volumeof interest.



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pressions and uprisings of formed surfaces. The spe-cial advantage of this innovative technology is theability to establish, for the first time in microscopy, adirect link between the microscope image and the 3Dmeasurement capability. There is no longer the needto struggle to find the area of interest in another de-vice, the image and 3D surface data coexist. Some ofthe many applications might be:

• Fracture analysis• Materials development• Drug discovery and production• Semiconductor• Life sciences research• MEMS • Entomology• Geology

Summary

Advances in image processing and computer tech-nology permit measurement of 3D surface parame-ters directly in stereomicroscopic images. Profile,area and even volumetric calculations can be per-formed in reconstructed 3D images. Therefore thespecimen does not need to be examined with other3D measurement devices; rather all analysis can beperformed with existing stereomicroscopes retrofit-ted with the new Leica Microsystems technology.This reduces investment costs while dramatically im-proving lab performance. All the measured values,the 3D models and other parameters can easily be ex-ported to standard files, integrated and furtherprocessed in reports. This new technology for stereo-microscopy by Leica Microsystems will become alaboratory standard for advanced research. > DG

Fig. 4 3D model of a solder paste with axesof coordinates visualised with Leica StereoExplorer

Fig. 5 Height-encoded phantom colourview of a crystal



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A capable and innovative partner in all aspects of 3D visualisation, LeicaMicrosystems has developed the world’s only 3D system for Leica Mstereomicroscopes which allows real 3D visualization without the needfor uncomfortable special glasses. The Leica 3D system is a profession-al, fully automatic solution for reconstruction, viewing, documentation,reporting, quantitative analysis and 3D measurement of three-dimen-sional surfaces and useful in all fields of science and technology. For vi-sualising and analysing specimen surfaces with an accuracy that, untilrecently, was nearly unthinkable, the innovative Leica 3D system nowmakes the process easier, more effective and more precise. In additionto imaging of surfaces, contact-free quantification of surface parame-ters such as height differences, roughness, surface areas or volumes isalso possible.

The three-dimensional image is created from a stereopair taken bythe Leica IC 3D digital stereo camera. The intuitive and user-friendlysoftware controls the Leica IC 3D camera and offers a variety of func-tions for optimally adjusting the imaging quality of the system to ob-tain a perfect 3D image. The Leica StereoExplorer software packageautomatically determines which pixels in the two slightly different im-ages belong together, and calculates the topography of the specimenas a surface model, taking into consideration the parameters of angleand magnification level. The 3D data record thus created serves asthe basis for surface and volume analyses.

The optional the high-resolution Leica ASD Auto-Stereoscopic DisplaySystem offers real time display of specimens in all three dimensions.Objects appear as concrete and vivid as in real life – no eyewear orother accessories are necessary. The depth of field and colour repro-duction correspond to the view through the eyepieces of a stereomi-croscope. > IH

The World’s Only Interactive Leica 3D System For Microscopy

Leica MacroFluo™ – The New Precision in Fluorescence Macroscopy

Leica Microsystems presents the world's only macrodocumentation systems for fluorescence methods,which have been developed in cooperation with theImaging Centre of IGBMC. Leica MacroFluo™ sys-tems with apochromatic manual or motorised 6.3:1 or16:1 zooms are the right choice for observing and di-gitally recording the effects of genetic defects inliving transgenic models the size of a whole mouse(zebrafish, Xenopus) with the highest precision andresolution in an intensely fluorescent field. TheMacroFluo™ concept combines the advantages ofmacroscopy – parallax-free imaging, generous fieldsof view and long working distances – with fluores-cence methods, including multicolour fluorescence,and high resolution. Unlike stereomicroscopy withtwo convergent optical paths, it guarantees an ab-solute maximum in precision when examining whole,living models, for measurements, and for digital im-age processing and analysis. That applies especiallyto processing with multifocus, overlay and deconvo-lution programs.

Leica MacroFluo™ systems deliver highest macro-scopic resolution for the experimental documentationof large objects: 340 Lp/mm with the 6.3:1 zoom and

336 Lp/mm with the 16:1 zoom (values with planapo-chromatic 1x objective and HC Plan wide-field eye-pieces 10x/25). With the planapochromatic 5x HR(high resolution) microscope objective, the wealth ofvisual information delivered by the MacroFluo™ sys-tems reaches the levels of traditional microscopy.Magnifications of up to 225x (6:1 zoom) or 575x (16:1zoom) can be achieved with a resolution of 1500Lp/mm. The bright, homogeneous fluorescence fieldautomatically adjusts to match the field of view whenzooming, guaranteeing precise illumination and maxi-mum light utilisation at any zoom position. The 5-posi-tion rapid filter changer supports the use of 26 differ-ent fluorescence filters for a wide range of fluores-cence techniques. The depth of field can be adjustedwith the integrated iris diaphragm.

Leica Microsystems offers a comprehensive range ofinnovative accessories for custom-tailored solutionsto meet any requirement or solve any problem. High-resolution FireWire cameras for fluorescence andsoftware (FW4000, IM1000 image overlay and multifo-cus) are available especially for documentation andimage analysis.

> IH


E V E N T S

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European Research Events

Exhibits:

Zebrafish – Genetics andDevelopmental BiologyJuly 13–16Dresden, GERMANY

BioScience 2005July 17–21Glasgow, UK

ELSOSeptember 3–6Dresden, GERMANY

MAF9September 4–7Lisbon, PORTUGAL

Transgenic Tech meetingSeptember 11–13Barcelona, SPAIN

Congress Nazionale di Neuroscienze 2005October 1–4Ischia, ITALY

Société Française de Biologiedu developpementOctober 1–4Obernai, FRANCE

Scanlab 2005October 11–13Stockholm, SWEDEN

BioTechnica 2005October 18–20Hannover, GERMANY

“Simply Brilliant” Workshops:

GERMANYCW 28: Kiel, BiocentreCW 36: BerlinCW 37: HamburgCW 39: TübingenCW 45: Berlin, Charité, Mitte

(Laser Microdissection Workshop)

For other countries please contactyour local Leica organisation.

Win a pair of Leica Binoculars, Leica Wristwatch or other great prizes!

Here is just a sampling of some of the events Leica Microsystems will participate inor organise in the 2nd half of 2005. For further information, please visit our website:www.leica-microsystems.com (click on events on the left)

CONTESTDear Customer:Please give us your comments on this first European Research Edition of reSOLUTIONmagazine. Send us your complete name and address of the institute where you work,along with your comments by going to the following link:

www.leica-microsystems.com/EU-Research

by August 31, 2005. Winners will be drawn on or around September 1, 2005 from all

completed entries.


http://www.leica-microsystems.com

http://www.leica-microsystems.com/EU-Research

re SEARCH 19

PAT H F I N D E R S O F T WA R E

Imprint

reSOLUTION is the magazine for Leica Microsystems customers and business friends

Publisher Leica Microsystems AG,Wetzlar (Germany)

EditorsPaul WismerRalf Niggemann

Editorial addressLeica Mikrosysteme Vertrieb GmbHLilienthalstraße 39 – 45DE-64625 BensheimTel.: +49 (0)6251-136-210Fax +49 (0)6251-136-155www.leica-microsystems.comE-Mail:[email protected]

Layout & ProductionCentral MarketingUwe NeumannLeica Microsystems AGErnst-Leitz-Strasse 17 – 37DE-35578 Wetzlar

Publishedtwice or three times a year

Cover PictureLeica Microsystems AGFluorescence Applications

Contributing editorsJohn Buckley > JBVerena Nickl > VNChristine Ludwig > CLIngrid Haack > IHDaniel Davis > DDDaniel Göggel > DG

New Cell Recognition Software

Pathfinder EditionTogether with the highly automated laser microdis-section system AS LMD, Leica now offers microdis-section with completely automated and integratedcell recognition. Collecting thousands of cells is nowpossible in just a few steps. The fully integrated soft-ware has a very powerful teaching mode to define therequired target cells by a few simple mouse clicks.

Top quality downstream kits fromQiagen – for reliable results

The Qiagen integrated solution for RT analysis givesLMD users ultra high sensitivity and specificity fordownstream analysis in RNA and DNA research. Nooptimisation is required. Optimised protocols are di-rectly available together with professional applica-tion support from Qiagen. The kits are specially de-signed for Leica Laser Microdissection.

Special 63x XT objective for Laser MicrodissectionDissecting single cells in the range of 5–20 µm in di-ameter requires special optics. With our 63x XT HCXPL FLUOTAR objective, these requirements are metwith brilliant image quality. The large working dis-tance of this objective allows easy and safe microdissection when working with groups or individual cells.

High quality service support for even longer than usual

Leica’s excellent worldwide service helps you to keepyour equipment running and up to date. With cuttingedge technologies like remote service via internet,support can be provided quickly and easily. The mod-ular system can easily be serviced by part exchange.The extended warranty will give you this performancefor one additional year.


Leica & Science!Famous scientists work with instruments from Leica Microsystems

Leading scientists who use instruments from Leica Microsystems value their high quality, technical perfection, technological

standards, service and large selection. When designing a new product, Leica Microsystems always puts the user first and

offers the best possible solution for each application and every budget: from routine microscope to high end systems for life

science and industry.

@www.leica-microsystems.com


reSolution_Research_News_No1

Documents

fluorescence imaging

x x x x x x0

high imaging quality

new leica af6000system

high end imaging systems

area of live cell imaging

high end imaging expertise

european scientists