Leica TCS LSI User Manual

Leica TCS LSI
The World‘s First Super Zoom Confocal!
From Gene to Cell - from Cell to Embryo
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The World‘s First Super Zoom Confocal
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Freedom for New Reserach
Easy to Achieve
- Large Scale Imaging
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An increasing number of scientists extend their focus of bio-research from single cell studies to entire organisms, analyzing the complex inter­action within whole animals. Thus, modern developmental biology is an emerging field of research, studying the dynamics of cell growth, differentiation processes and the development of organs
As living organisms grow in three dimensions these studies require imaging systems which pro­vide - beside high resolution – a large workspace and field of view. Leica Microsystems introduces a pioneering imaging system for developmental
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biology which provides all these features in one: the new Leica TCS LSI.
in vivo
.
Leica TCS LSI
The world‘s first super zoom confocal!
Leica TCS LSI is the first super zoom 3D-confocal, offering high resolution plus large field of view for
in vivo
(LSI) platform provides generous workspace and adapts perfectly to the experiment needs of native specimen analysis.
True confocal technology is used to provide crys­tal clear images of highest spectral resolution, revealing finest details of the model organism, no matter if drosophila fly, mouse, plant or zebra fish.
An automated optical zoom system allows for seamless magnification change on demand, easy switching from overview to details and free 3D navigation through the specimen.
imaging. The new Large Scale Imaging
Gain new insights with TCS LSI and study processes of life – from egg to fly, from embryo to adult.
in vivo
imaging by Leica
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Benefit from freedom for new applications offered by Leica TCS LSI, feel free to zoom in and out and travel time resolved in all di­mensions
in vivo
through your specimen.
Leica TCS LSI
New perspectives of life: From cell to animal
A system tailored to your needs
With the Leica TCS LSI, study the process of life from embryo to adult. Due to the functional and workflow oriented design, even sample preparation and orientation can be directly performed on the imaging system.
Take advantage of the combination of high-resolution confocal technology and large field macro zoom imaging. A generous workspace provides extended freedom for Zoom in and out, from overview to area of interest at highest reso­lution and obtain fascinating results.
in vivo
experiments.
Features
Zoom factor up to 16x
Field of view up to 16 mm
Generous workspace
Free sample access by wing door
Easy sample manipulation
Largest motorized z-control
Large xy-travel range for optimal
positioning
Precise z-control by galvo stage
From cell to embryo
Leica TCS LSI enables you to visualize cell growth and the fasci­nating differentiation of cells into organs in real life from cell to embryo – in one system!
Identify new pathways from gene to cell, from cell to animal; or ex­amine the influence of genomic defects on the whole animal. Study time resolved 4D processes at highest resolution easily as Advanced Time Lapse software is provided for in vivo studies.
Analyze protein interactions or test the influence of drugs in bio-medical research.
Visualize the development of life: the Leica TCS LSI Large Scale Imaging System provides highest image quality, easy operation and maximum flexibility.
in vivo
Largest workspace...
...for finest insights
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Leica TCS LSI – The Large Scale Ima
High Resolution in 2D and 3D
From micro to macro ...
Get the big picture of large specimen in 2D or 3D. Use the Leica LAS AF 3D-visualisation package for reconstruction of Drosophila larvae. (Left)
... zoom in seamlessly into the finest details!
The motorized optical zoom offers the advantage of flexible magnification to identify the finest details of your model organism. Backbone details of zebra fish or eye of the
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drosophila larvae, both in highest resolution. (Bottom)
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4D-Advanced Time Tapse
From larvae to fly ...
Tracking the development of life over time offers exciting insights for embryology and morphogenesis – all in the same specimen! (Right)
... from egg to fish, achieve excellent results in 4D!
To obtain exciting views of the development of organs with LAS AF Live Data Mode software and see the backbone for­mation during the growth of a zebra fish. (Bottom)
T= 0 h
T= 10 h
T= 7 h
T= 15 h
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Dynamic studies
2D, 3D and 4D analysis
Advanced Time Lapse
Spectral analysis
Photo activation with 405 nm
Micromanipulation
in vivo
More freedom than ever: For various specimen sizes and research questions, micro- and macro objectives can be used. High resolution and largest field of view enable a wide range of new applications. For excellent new research opportunities the Leica TCS LSI offers variable magnification with a field of view up to 16 mm plus large and precise z-positioning – all in one system.
In Vivo
Large Scale Imaging
High resolution from micro to macro
A comfortable start
Instead of engineering the living object through holes into incuba­tors, start to place your specimen easy and securely through wide-open wing doors of the laser safety chamber. Gain overview first by digital camera for easy orientation. Fine-focus precisely either with the tuning wheel of Universal Microscope Control or by LAS AF software.
Study the changes of life in 3D
With Leica TCS LSI, you navigate through your model organism from large to small and back, identifying functions and obtaining insights never seen before. From the entire animal up to the finest detail the motorized zoom offers the advantage to select any area of interest – without changing the objective! Fine-tune the z-posi­tion through the highly precise SuperZ Galvo stage and orientate the sample perfectly in xy-direction with a high precision motor­ized stage.
Ultra dynamic z-control
The flexibility of the Leica TCS LSI is unique in confocal mi­croscopy. The SuperZ Galvo stage offers backlash free sensitive vertical positioning with a maximum travel range of 1500 µm. The fine focus integrated in the motorized zoom system allows further to extend the focus range by 10 mm. Finally, ultimate z-position con­trol is achieved by the motor focus itself, travelling up to 150 mm.
Seamless zooming in and out
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Free xy-sample positioning
Leica TCS LSI systems provide a maximum travel range, indepen­dent of the type of stage. Both, manual and automated stages of­fer a wide range for ideal sample positioning.
Increased cell-viability and highest resolution
Obtain brilliant fluorescent images over long time. Ultimate image quality is provided by true confocal point scanning technology. The Leica TCS LSI offers a variety of automated tools to adjust ex­citation and emission perfectly to your individual sample condi­tions. Maximize signal efficiency with the freely tunable spectral detector, the highly dynamic photomultiplier and minimize laser excitation power with the 100%-tuneable AOTF-attenuation. This maximizes the lifetime of your specimen.
Developmental
Biology
Plant Research
Growth of
Animal
Structure of
the Leaf
Studies of the
Anatomy
I S L S C T a c i e L
Crop Design
Research areas
Developmental biology
Embryology studies
Morphogenesis
Embryo genetics
Plant science
Genetics
Proteomics
Neurology
Applications
Cancer research
Agriculture investigations
Pharmaco screening
Seed development
Cell development
Heart diseases
Brain development
Genetics
Neurology
Examples of Leica TCS LSI applications
Inuence of
new Drugs
Structure of
the Brain
Lead Finding
Research
Neuro-
Research in
degenerative
Diseases of the
Diseases
Elderly
Field of view depending on objective and zoom adjustment
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New Dimensions
Generous workspace
Free sample access by wing doors
Easy sample manipulation
Largest motorized z-control
Hardware for most flexible use
Large xy-travel range for optimal
positioning
Precise z-control by galvo stage
Motorized and manual xy-stages
Accessories for environmental
control of temperature, CO2,
humidity
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Making life visible in 4D
Even more than the static view, it is fascinating to observe the devel­opment of whole organisms over time. The Leica TCS LSI with LAS AF Live Data Mode Software offers perfect automation for cell de­velopment studies, from egg to embryo. Individual experiments can be easily combined to a fully automated workflow. The door is open for continuous studies from cell to adult.
Advanced Time Lapse
High resolution from cell to embryo
Perfect climate included
Optimal growth conditions for living animals can be provided by a wide range of accessories. The laser safety cabinet is converted in­to a climate chamber just by adopting a heating unit for precise tem­perature control. Stage adapters for CO fer optimal sample conditions. Even during the experiment, remote controlled manipulators enable active specimen handling within the native environment.
-gas and humidification of-
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All accessories are offered in modular and approved kits for easy system upgrade on demand, ready for todays and future experi­ments.
Following the changes of life
Studying movements of cancer cells in bio-medical research, inves­tigating translocations after photo-activation, studying the growth of bio films on large implants: The creativity for future experiments will come from you, the extended freedom for new research applica­tions is provided by Leica TCS LSI.
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Zebra fish development video
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Confocal technology
Highest resolution
True confocal point scanner
Spectral detector
Solid state laser
405, 488, 532, 635 nm excitation
AOTF controlled
Fully automated
Easy to use
Crystal clear 3D images – The benefits of true spectral confocal
imaging
Leica TCS LSI is equipped with a true spectral confocal scanner that provides ultimate crisp resolution. Scanning the specimen in thin op­tical layers and detecting the fluorescence signal point-by-point re­sults in images free from the stray light of adjacent elements. The re­sult – brilliant images at very high resolution.
The World’s First Super
Zoom Confocal
Technique for new perspectives
Beam Splitter
The Confocal Principle
Detector
Confocal pinhole
Laser
Objective
Focal plane
The Spectral Principle
Information from each signal in all optical sections is reconstructed by intelligent software into excellent 3D images, resolving the smallest detail of the specimen's structure. To ensure high quality imaging, optimal excitation is provided by up to four solid-state lasers with 488, 532 and 635 nm lines for common dyes. The broad range of applications is extended by the 405 nm laser option for nuclear staining.
Maximum signal efficiency by spectral detection
Maximize the signal independent from barriers posed by fixed fil­ters and tune freely the emission band from 430 nm to 750 nm. Cross-talk of different dyes in one sample is prevented when tun­ing the spectral detector precisely in 10 nm steps to the respective dye. For unrivalled detection efficiency, the Leica TCS LSI uses a
Light beam
Prism
Variable Spectral Detector
Collimation Optics
Detector
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prism spectral detector. Minimize bleaching and cell damage by op­timizing the detection range and reduce the excitation power via Acousto-Optical Tuneable Filter (AOTF).
Highly innovative
Zoom freely in and out, switching between overview and smallest detail without changing the objectives. The Leica Z16 APO A 16:1 su­per zoom offers the largest magnification range from 0,57x to 9,2x, whereas the Z6 APO A ranges from 0,57x to 3.6x – at excellent opti­cal performance. The Z-zoom systems are fully apochromatic and allow to adjust magnification continuously and parallax-free.
Benefit from the motorized versions Z16 APO A and Z6 APO A: Fully software controlled, the magnification can be completely altered without touching the imaging system. To achieve precise focusing for sharpest images over a wide range of 10 mm, all motorized zooms are equipped additionally with a motorized fine focus lens.
Macro and Micro: New dimensions for largest specimen
Benefit from the high quality macro objectives of Leica Microsys­tems. A novelty in confocal imaging: the tremendous working dis­tance (WD) of 97 mm and a field of view (FOV) of 16 mm provided by the 1x apochromatic macro objective. Additionally, classic micro­objectives can be adapted to use the Leica TCS LSI as a classical confocal. With high resolution, high numerical aperture lens sys­tems, finest details at maximum resolution become visible.
New zoom optics
Change magnification by fingertip
Continuously variable magnification
by 16x
Motorized
No objective change required
Apochromatic, parallax-free optics
Macro and micro objectives
1x, 2x, 5x, 10x, 40x, 63x
Adjusting of magnification by motorized zoom
Intensity (arbitrary units)
Wavelenght (nm)
Sequential Scan
λ-Scan
Intensity (arbitrary units)
Wavelenght (nm)
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Highest Flexibility
Dynamic confocal imaging
Spectral band tuning
0 – 100% AOTF laser control
Variable pinhole
Variable magnification
Motorized z-zooms
Full objective band from 1x to 63x
FOV max: 16 mm
WD max: 97 mm
Z-range max: 150 mm
Observation and handling of large specimen becomes facile: Over­come the limits of fixed magnifications, small fields of view and low working distances.
Freedom for New Research
Enter new dimensions for flexible applications
The new Leica TCS LSI offers unrivaled flexibility for high resolution imaging. Enjoy the freedom and navigate through your model organ­ism in all three dimensions.
Stage inserts for various applications
Micro­Lenses
A system flexible for all experiment needs
Observe any specimen straightforward: The large workspace al­lows for studying even an entire mouse. Select the objective which corresponds best to your experiment. Profit from the fundamentally new design of the laser safety chamber and modify your sample ac­tively by adding a drug to the test population, as even manipulators fit in easily. Through the wide-open wing doors, specimen insertion is facile and handling is comfortable.
Open the door to new applications
Leica TCS LSI is the true first high resolution imaging system offering this extraordinary flexibility. Instead of switching between different instruments, you can do your research and design new experiments with one system. From micro to macro, Leica TCS LSI opens up fu­ture for ultimate new research topics.
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Easy to use software and workflow-oriented hardware minimizes the training effort and allows scientists to work with the Leica TCS LSI confocal straight away.
Easy to Achieve
Efficient operation for faster success
Easy interfacing: requires less training and enables faster working
From magnification up to z-position, remote control the instrument easily. Minimum training is required even for newcomers. The standardized intuitive user interface of Leica LAS AF enables you to start your work autonomously from the first button click.
Ergonomic software with workflow-oriented screens guides you through your experiment from the first confocal image up to the full 3D reconstruction of your specimen. The effect: Highly efficient imaging and best results on the first shot.
Minimal training effort
Large workspace
Variable magnification
Workflow orientated
Fully automated system
Comfortable operation
Easy sample access and
manipulation
Compact design
Additional functionality like deconvolution or spectral unmixing en­sure application flexibility for the future.
Easy sample handling
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Efficient Operation for
Faster Success
Workflow orientated hardware design
The time for preparation, pre-selection and orientation of the speci­men reduces enormously as macro and high-resolution confocal are combined in one in vivo system.
Saving costs and lab space
The Leica TCS LSI combines both, a macro and a confocal imaging system. The provision of various imaging tools becomes obsolete.
Less stress for the specimen
By avoiding the transportation between different imaging tools, stress to living specimen is reduced, the survival rate increased.
Easy confocal imaging:
6 Steps to 3D
1. Start the system
2. Insert specimen, align focus
3. Select instrument settings
4. Define z-range and acquire
5. Calculate 3D image
6. Save and close
Comfortable handling
Profit from workflow orientated design of the Leica TCS LSI: speci­men exchange can be performed fast and secure through wide open wing doors.
Minimal pre-processing
As all sizes fit easily into one system test organisms by avoiding histopathological tissue processing.
Efficiency for research
Enjoy the efficient operation of the fully automated system: Change magnification quickly on demand and avoid the possible risks during objective changes.
Fast results
Save the time for post processing algorithms as the Leica TCS LSI provides excellent true confocal raw data immediately from your experiment.
in vivo
, reduce the number of
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Less specimen needed
With Leica TCS LSI, more experiments now can be performed with a single specimen to ensure highest reproducibility and optimal results.
Time Saving
Operate the Leica TCS LSI where your research equipment is, shorten the lane and place the instrument directly in your lab envi­ronment.
No need for special room conditions
The dark acrylic glass protects the experiment against light and as the system works at room temperature, additional costs for special room conditions are not required.
A robust system
Stable technology and long lifetime components make the Leica TCS LSI robust and minimize maintenance. Predefined upgrade kits offer extensions for future research.
Ergonomic software
Common Leica LAS AF platform
Ergonomic and standardized user
interface
Predefined settings
Automated long term observation
with Advanced Time Lapse
Easy data transfer
Analyse specimen with up to eight
colors
Efficient operation and excellent results
Brilliant images in short time with less test organisms ensure fast success.
Prepared for the future
Achieve excellent results from micro to macro today and inves­tigate ultimate new research topics in the future with the new Leica TCS LSI.
Leica TSC LSI was developed in co­operation with Jean Luc Vonesch and Didier Hentsch of the Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Illkirch, France.
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1 Confocal scanhead
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Supply unit
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Laser safety chamber
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Wing doors
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Z-zoom, motorized
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SuperZ Galvo stage
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Motor focus drive
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DFC camera option
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Anti-vibration table, passive
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Monitors
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Mouse
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Keyboard
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UMC control
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XY-stage control
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Heat pipe adapter
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EL6000 fluorescence llumination
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Computer table
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Leica Design by Christophe Apothéloz
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Acknowledgements:
1. Mouse transgenic embryo
E10.5 mouse transgenic embryo: EpaxialMyf5 eGFP; immuno-stained for GFP-Alexa 488; the embryonic muscle fibers and the heart are stained with Desmin-Cy3. From top to bottom around 3.5 mm. Courtesy of: Aurélie Jory and Shahragim Tajbakhsh, Cellules Souches et Développement, Institut Pasteur, Paris, France and Imaging centre of IGBMC, IGBMC, Illkirch, France.
2. Drosophila melanogaster, head of larvae
Head with nerve system and photoreceptors, Red: Cy3, Green: Alexa 488.
3, 13. Lilly of the valley, rhizome
Convallaria sp.: Rhizome with concentric vascular bundles. Red: cell wall, Green: chloroplasts.
4. Mouse cerebellum
Mus musculus. Cerebellum of mouse P21. Green: Alexa 488, antibodies against GFAP, glial marker, Blue: DAPI. Courtesy of: Giovanni Marchetti, Team E. Georges-Labouesse, Imaging centre of IGBMC, IGBMC, Illkirch, France.
5. Mouse transgenic embryo
Mus musculus. E10.5 mouse transgenic embryo: EpaxialMyf5 eGFP; immuno-stained for GFP-Alexa 488; embryonic muscle fibers and heart stained with Desmin-Cy3. Size from top to bottom: 3.5 mm. Top: RGB-channels. Courtesy of: Aurélie Jory, Cellules Souches et Développement, Institut Pasteur, Paris, France.
6. Chicken, spinal cord
Gallus gallus. Dorsal view of a whole chicken embryo at 10-12 somite-stage, 6h post-electroporation using two DNA constructs: CMV-GFP and neural specific enhancer β−Galactosidase reporter gene. Courtesy of: Isabelle Le Roux and Shahragim Tajbakhsh, Cellules Souches et Développement, Institut Pasteur, Paris, France, Imaging centre of IGBMC, Illkirch, France.
7.
Drosophila melanogaster
Drosophila melanogaster
Ventral view, larvae L1.
8, 9. Mouse transgenic embryo, interlimb somites
Five interlimb somites of an E10.5 mouse transgenic embryo: EpaxialMyf5 eGFP; immuno-stained for GFP-Alexa 488; embryonic muscle fibers stained with Desmin-Cy3, the nuclei are revealed with Hoechst Size from top to bottom: 3.5 mm left, 800 µm right. Courtesy of: Aurélie Jory and Shahragim Tajbakhsh, Cellules Souches et Développement, Institut Pasteur, Paris, France and Imaging centre of IGBMC, IGBMC, Illkirch, France.
10.
Drosophila melanogaster
Larvae, Head with nerve system, Red: Cy3, Green: Alexa 488, Eye: Photoreceptors with nerve system. Red: Cy3.
12. Zebra fish, novocord development
Danio Rerio. Red: Rhodamine-dextran, Green: GFP, labelling of the notochord. Courtesy of Sophie Dal­Pra, Team B&C Thisse, Imaging centre of IGBMC, IGBMC, Illkirch, France
14. Mouse cerebellum
Mus musculus. Cerebellum of mouse P21. Violet: Alexa488, antibodies against GFAP, glial marker, Blue: DAPI. Courtesy of: Giovanni Marchetti, Team E. Georges-Labouesse, Imaging centre of IGBMC, IGBMC, Illkirch, France
15. Zebra fish
Danio Rerio. Red: Cy3, z-disk staining. Blue: DAPI, nucleoli.
, central nerve system
. Green: GFP. Transgenic fluorescent protein fluorescence in larval CNS.
, head and eye.
www.leica-microsystems.com/tcs_lsi
16. Zebra fish development
Danio Rerio. Embryo, T=0h, 6h, 11h, 17h. Bodipy TR. Courtesy of: Jabier Gallego Llamas, Team P. Dolle, Imaging centre of IGBMC, IGBMC, Illkirch, France.
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Drosophila melanogaster
Larvae Blue: DAPI, nucleoli, Green: Alexa 488, CNS.
, nerve system development.
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