Workshops - How to Sign Up

Please see attached Excel spreadsheet for workshop titles and timings.Available workshop time slots are indicated by the blue boxes (not white).Please select the workshops you wish to attend by marking an ‘x’ in the appropriate blue box time slot.Many workshops are repeated to give you several opportunities to attend.Please make sure that you only have one ‘x’ in any column as workshops will be running in parallel.

If you are attending any ‘b’ workshops you may not be able to attend one of the numbered workshop either side of the ‘b’.

The LaVision workshop will be held off-site at the Beatson Institute. Some time will be lost in the transportation back and forth between sites. So for example, If you select workshop 1B (pink Box) you may select workshop 3( pink Box) but not workshop 1 ( yellow box ). If you select workshop 2B ( yellow box) you may also select workshop 1 ( yellow box) but not workshop 3 ( pink Box).
Similar rules apply on Thursday.

Delegates will be allocated to workshops on a first come first served basis. Please be aware that numbers are strictly limited for each workshop. If any of your selections are not possible you will be contacted with an alternative time slot.

Please complete your selection, put your name at the top where specified and return a copy of this spreadsheet to Victoria Lee by email, Victoria@rms.org.uk, as soon as possible.

The deadline for early registration to the workshops is Wednesday 3^rd June at 12.00. If you do not register by this date, you will be required to register for the Workshops at the ELMI Meeting, and availability may be very limited. We cannot guarantee you will be able to attend the workshop of your choice if you do not pre-register.

ELMI 2009 Workshops will be provided by the following companies: Click on logo for workshop Abstract.

ELMI 2009 exhibition stands will be provided by the following companies:

Michael Sommerauer

AHF analysentechnik AG, Kohlplattenweg 18, D-72074 Tübingen

Perfectly matched optical filters are essential tools to image samples in fluorescence analysis.Sputtered optical filters show up to 99% transmission, steepest edges, excellent blocking and high environmental stability. Single-or multiband filters - also for many laser lines - are available.

 During the workshop we will discuss filter setups for applications-,multicolor analysis,time lapse experiments,single molecule detection (SMD), TIRF, multiphoton microscopy, SHG,  RAMAN / CARS

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Workshop 2
 

Optical Filters – Specifying filters for advanced microscopy

Michael Sommerauer

AHF analysentechnik AG, Kohlplattenweg 18, D-72074 Tübingen

Advanced microscopy requires very sophisticated optical filter setups. Besides optical specifications also other physical characteristics are decisive for successful experiments.

We will discuss flatness of optical components, wavefront distortion, dispersion and effects of polarized light.

Filters can be stacked or used under angles of incidence ≠ 0 degrees or cone angles ≠ 0 degrees.  Options and limitations will be discussed.

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Bitplane - 3D and 4D Image Visualization & Analysis Workshop

Running the workshops:

Luciano Lucas, luciano@bitplane.com        Dieter Goehlmann, eusupport@bitplane.com

Shaun Fitch, shaun@bitplane.com 

In the Bitplane workshop you will enter the “Champions League” for 3D and 4D Image Visualization & Analysis. The recently updated version, IMARIS 6.3, is one of the most advanced software tools for researchers whose daily business includes microscopy. With its seven modules, IMARIS enables top researchers to easily, load, visualize and analyze data from a few megabytes to more than 50 gigabytes in size, from almost all microscope image formats. 

The workshops will introduce you to the IMARIS software, including all the new features, and will enable you to see how you can obtain stunning results from your datasets.

Come and explore the third dimension!

IMARIS6.3: http://www.bitplane.com/

IMARIS modules: http://www.bitplane.com/go/products

IMARIS movies: http://www.bitplane.com/go/web-training

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European Light Microscopy Initiative – Nikon UK 

Confocal is A1ive

Powerful and versatile, Nikon’s new generation A1 Confocal systems (A1, A1R and A1Si) incorporates novel optical and electronic technologies to provide unrivalled speed, sensitivity and flexibility in live cell imaging and without undue cell stress.  Nikon will be demonstrating the use of fast resonant scanners in combination with standard galvo-based scanners and the use of a second pair of scanners for simultaneous bleaching (FRAP). 

New Macro-Confocal Imaging

Nikon will be launching its new Macro-Confocal imaging system during ELMI 2009.  This new Macro-Confocal allows the researcher to image large specimens in confocal mode using tele-centric optics, making it ideal for application areas such as developmental biology, cell biology, stem cell, tissue research and many more.  The Nikon Macro-Confocal system can be equipped with true spectral imaging and controlled light excitation microscopy (CLEM).  Both techniques will be demonstrated during the workshops. 

Evanescent Wave Imaging Perfected

Total Internal Reflection Fluorescence (TIRF) microscopy is a surface imaging technique that can be used to obtain a very thin optical section of a specimen whilst minimising background noise.  TIRF is a highly sensitive widefield camera-based technique providing an excellent signal-to-noise ratio compared with epi fluorescence or confocal imaging methods.   Coupled with Nikon’s Eclipse Ti inverted microscope and outstanding optics, Nikon will show how its Perfect Focus System will revolutionise your TIRF applications.     

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LaVision

Expand Single-beam 2-photon microscopy with Multibeam Technology for higher resolution and higher speed. Add OPO for double-the-depth tissue and intravital imaging.

Two-photon microscopy is indispensable for deep tissue- and intravital-imaging. However, current technology based on single-beam point-scanning has reached sensitivity- and speed-limits as higher performance requires higher laser-power leading to sample-degradation. LaVision BioTec’s TriM Scope utilize a single beam scanner as well as a multi-focal scanhead, splitting a laser-beam into a line of 64 foci allowing sample illumination in real-time at full laser-power. This technology requires CCD-field-detection contrary to conventional detection by photomultipliers (PMT). The advantage of PMT-detection starts at imaging larger depths due to a better signal-to-noise-ratio, while CCDs allow real-time detection of rapid processes. LaVision BioTec’s TriM Scope provides both techniques, which can be compared during the workshop.

A new technology expanding the excitation range beyond 1000 nm will be presented. The optical parametric oscillator OPO produces fs IR light at high repetition rate. The IR light penetrates even deeper into tissue and excites new red fluorescent proteins (RFP, DSRed etc.) at their true excitation wavelenght. Expect to double the depth you can reach!

Finally, we will discuss our latest addition: a sensitive and fast TCSPC FLIM detector. Until now one has had to choose between fast but rather shallow imaging, and deep but slow. For the first time FLIM imaging deep into a tissue can be realised at biologically relevant speeds.

LVBT ELMI  Presentation

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Advanced Quantitative Low Light CCD and EMCCD Imaging for Bio-Research

Jim Schumacher & Deepak K. Sharma

Photometrics, 3440 East Britannia Drive, Tucson, AZ, USA 85745

Keywords: EMCCD, on-chip gain, CCD imaging, incident photons, Quant-View, Rapid-Cal

 Photometrics will present a simple yet comprehensive course on measurement of photons via CCD imaging. We will start with a short talk explaining basic concepts of CCD imaging and fundamentals of CCD operation. Concepts such as speed, binning, linearity, read noise, system gain and calculation of incident photons will be covered. Lab handouts will be given and short lab tasks will be performed. These will include understanding and consulting the specifications provided by CCD camera manufacturers on their data sheets. An exercise will be performed where participants will measure a mean variance curve which will be used to calculate photo-electrons per grey level.  User will then be able to back calculate their imaging data into photo-electrons and even probable number of incident photons. Advanced quantitative EMCCD imaging will also be demonstrated utilizing propriety advanced EM-CCD technology such as Rapid-Cal and Quant-View and the importance of such quantitative low light measurements in bio research imaging will also be discussed.

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ELMI 2009 Conference
Glasgow, UK

Presenter:  Paul C. Goodwin
Technical Fellow and Sr. Product Line Manager
Applied Precision, Inc.

Title:  Super Resolution and High-speed Imaging with DeltaVision OMX

Abstract:

In this workshop we will describe the DeltaVision OMX system from Applied Precision. This system, based on the OMX Microscope developed Dr. John Sedat and colleagues at UCSF, is capable of fast, live-cell imaging for fast dynamic events and is also capable of super-resolution imaging using structured illumination methods per Mats Gustafsson and others (3D-SIM). With 3D-SIM, the DeltaVision OMX system is routinely achieving ~100nm lateral and ~250nm axial resolution with multiple probes well away from the cover glass. In this workshop we will also provide instruction into the mechanics of 3D-SIM, image reconstruction methods, and hands on post-processing of 3D-SIM images. Finally, a number of examples will be given where the 3D-SIM is being used in biomedical research.

Note: Due to the size of the actual DeltaVision OMX system, the acquisition system will not be present at ELMI.

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Carl Zeiss Workshop 1

CONFOCAL MICROSCOPY, IMPROVEMENTS IN SIGNAL TO NOISE AND SPECTRAL IMAGING

 To use the full capacity of various fluorescent dyes and labelling techniques it is essential to clearly identify and separate their emission signals. This is often complicated by similar excitation and emission spectra. As a consequence, conventional imaging approaches with limited flexibility in detection and excitation do not provide an accurate identification of fluorescent components with similar spectral properties.

The innovative changes in the new design of the universal TwinGate beamsplitter and the QUASAR Detector in the LSM 710 take spectral imaging to a new level. The new InTune laser with free selectable excitation wavelength allows an exact fine tuning of imaging strategies. On the detection side, the arrangement of a 34 channel detection allows fast spectral detection with simultaneous acquisition and ideal SNR. 

The excellent sensitivity of this calibrated and linearized detector unit (the QUASAR detector) and its outstanding suppression of noise and excitation laser light provides the basis for advanced processing techniques. Advanced algorithms such as weighted linear unmixing greatly enhance the accuracy of the unmixing result. Overall, the linear unmixing technology coming with the LSM 710 results in improved precision and signal strength of crosstalk-free images. 

For multiphoton imaging, the detection of the emission signals needs to be as efficient as possible. The new LSM 710 NLO from Carl Zeiss offers a cascade of up to 5 PMT-based NDDs and a single detector using the GaAsP technology. The very low dark noise of this detector produces outstanding images with very high signal to noise ratios.

With the new ZEN 2009 software, all these advantages are available now in a 64-bit software environment for better RAM and computing performance.

Carl Zeiss Workshop 2

Recent technological advancements in Confocal Microscopy
From a specialists’ system to the high end microscope for all – the LSM 700

Laser scanning microscopes (LSMs) together with an increasing number of fluorochromes provide a powerful tool to directly visualize the complex relationships and interactions of cell components and molecules in their natural context. By eliminating out of focus fluorescent signal the LSMs produce optical sections and provide a clear image of the sample without having to physically harm the sample.

The new LSM 700 from Carl Zeiss MicroImaging pushes sensitivity, image quality and flexibility to next level, by the means of the innovative changes in beampath design with its highly corrected optics and the outstanding electronics of the PMT detectors.

Up to four stable solid-state lasers are individually guided and color-corrected into the optical axis of the system, 

Efficient separation of the fluorescence signals is mastered by the unique variable secondary dichroic (VSD), granting flexible adjustments to the image setup for presently and future used fluorochromes ensuring unambiguous results. The innovative integration of the VSD beamsplitter into the LSM 700 concept provides a highly efficient method of spectral image acquisition, enabling for precise separation of even highly overlapping fluorescent signals.  

Above and beyond its superb imaging capabilities, the LSM 700 provides the means to conduct complex multidimensional experiments, such as 3D, live cell imaging, ion imaging, as well as photomanipulation setups as used for example in FRAP, FLIP and photoactivation.

The intuitive ZEN software provides easy control of all technical features of the LSM 700. The integrated Smart Setup tool conveniently assists you to set the microscope for optimum image acquisition within the shortest possible time.

Carl Zeiss Workshop 3

Cell ObserverÒ SD – the Solution for Fast Confocal Imaging on Living Cells

Carl Zeiss combines HighEnd-Technologies: ms-precision of Cell ObserverÒ, the live cell imaging platform from Carl Zeiss, with high speed optical sectioning of CSU-X1, the latest confocal scanning unit from Yokogawa Electric Corporation (Japan). In combination, Cell Observer SD offers fast confocal image acquisition, precise timing and highest flexibility.

The modular concept of Cell ObserverÒ allows configuration of an individual system to suit your applicational needs. A range of high magnification objectives especially designed for live cell imaging  and perfectly suited for use with CSU-X1; Cameras offering highest sensitivity and acquisition speed and a variety of incubation solutions. All components are fully integrated into AxioVision, the software from Carl Zeiss allowing convenient control of complex experiments and documentation of all imaging parameters including environmental parameters like temperature or CO2. A high speed acquisition mode with support for simultaneous read-out of two cameras allows optimum use of all features offered by the CSU-X1.

The latest model of the renowned microlense-enhanced spinnig disc systems from Yokogawa offers frame rates only limited by camera speed, increased light efficiency, a high speed filter wheel for fast multicolour experiments and a dual camera option for simultaneous detection of two emission channels.

Cell ObserverÒ SD with its full integration of CSU-X1 into AxioVision is the perfect system for fast confocal time lapse imaging of living cells with lowest phototoxicity and highest precision. Cell Observer SD is a complete system from one partner with full service and support from Carl Zeiss.

Carl Zeiss Workshop 4

An analog approach goes digital : New developments in TIRF microscopy

Over the past twenty years, total internal reflection fluorescence (TIRF) microscopy has been established as a powerful method to observe and analyze dynamic processes around the cellular membrane. This holds also for in vitro experiments.

The big advantage of the TIRF microscopy over other imaging techniques derives from its principle. The sample itself is not directly illuminated by fluorescence light, but only molecules that are located within an evanescent field can be excited. Because of the preliminaries of the technique, namely high numerical aperture lenses and life cell labels, the use of TIRF microscopy could only be recognized by a broader audience, after other powerful tools like fluorescent proteins and other dyes and markers have been established in the biosciences.

With modern microscopes and digital imaging it is know possible for the first time to even combine TIRF microscopy with other applications, e.g. electro physiological approaches or other image forming methods like atomic force microscopy, or even with laser manipulation techniques.

The combination of imaging data with data from force measurements or electrophysiology will lead to new insights in cell biology and biophysics in the near future.

In this talk several options and approaches to the use of TIRF microscopy and possible combinations with other techniques will be discussed.

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OMERO Workshop

Chris Allan

The OMERO Workshop will be a general introduction to the OMERO Platform,
focusing on the client tools OMERO.importer, OMERO.insight,
OMERO.editor, OMERO.ij and Bio-Formats.

We will describe the type of client-server application OMERO is and
provide an introduction to OMERO.importer. We encourage people to bring
some of their own original microscopy data files (e.g. LSM, Leica,
DeltaVision, Zeiss, ...) on a USB key, hard disk or DVD. We will also
have sample data available. OMERO.insight will then be introduced and
participants will be shown how to manage, annotate, visualise and
analyse their images. The metadata captured during the import process
will be shown and additional experimental metadata will be explored
using OMERO.editor. Integration with ImageJ will be briefly shown using
the Bio-Formats and OMERO.ij plugins.
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imageJ

Daniel James

WhiteFiji is an image processing package based on ImageJ.For users, Fiji is easy to install and update, bundles a set of plugins in a coherent menu structure (and updatable), along with comprehensive documentation.
For developers, Fiji as an open source project is hosted on a git source version control repository, with access to the source code of all internals, libraries and plugins, and eases the development and scripting of plugins.