Leica AF6000E – AF7000
Advanced Widefield Systems
Living up to Life
Live Cell Imaging Solutions
That Grow with Research Needs
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Flexibility and Modularity
– Individually optimized system configurations
– Available for upright, inverted and stereo
microscopes
– A range of application wizards, process
and quantification tools
– A wide choice of hardware components
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Upgradability
– Systems that grow with changing
research needs
– From entry-level systems to high-end
workstations for real-time studies
– Convenient, on site upgrade process
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Integration
– Perfect harmony between hardware
and software
– Maximum system reliability
– Easy user handling
|||
One Platform
– One software platform for all widefield
research and confocal microscope systems
– Ideal for imaging centers and
multi-user facilities
– Minimum training required
2
Biological and medical researchers are probing deeper into the
secrets of life, and widefield fluorescence microscopy is a
key technology for imaging the living cell. The developmental
and molecular processes of the cell can be visualized directly
by widefield fluorescence techniques to shed new light on the
mysteries of life. Leica Microsystems’ new Advanced Widefield Systems provide the leading innovations to advance
your own research quest and unlock the secrets of life.
Users all over the world have benefited from the speed, reliability, brilliant image quality and ease of use of the fluorescence research systems from Leica Microsystems for many
years. Together with leading scientists, Leica Microsystems
has further developed its successful fluorescence solutions
into a modular Leica Advanced Widefield System line that
can be customized to suit all live cell imaging requirements.
The Leica Advanced Widefield System Series offers the right
tool for your research; from routine imaging and documentation
to live cell imaging workstations for complex real-time experiments. Regardless of the application, all Leica Advanced Widefield Systems utilize the same easy-to-use software platform,
which is shared with the Leica Microsystems confocal systems.
The Leica Advanced Widefield System Line consists of four modular
product solutions, which can be customized for different levels of
application. The modular design affords you the freedom to grow
the system and to choose hardware and software components as
needed.
❚ Leica AF6000 E – The entry-level software solution for high-quality
fluorescence imaging and documentation. Compatible with a wide
range of manual and automated microscopes and cameras from
Leica Microsystems.
❚ Leica AF6000 – The flexible system for a variety of applications in
fluorescence microscopy and image analysis including time-lapse
experiments, multi-positioning, z-stacking and deconvolution.
❚ Leica AF6500 – The high-speed fluorescence system with all the
functionality of the Leica AF6000 plus real-time controlled image
acquisition for ultra fast 2D time-lapse experiments, ratio imaging,
and triggering of peripheral hardware components.
Dendritic cells infected with bacteria
Green: GFP expressing enteropathogenic E. coli
Red membrane staining: cy3 labeled anti cd44 antibody
Courtesy of: Nadzeya Kramko, ZMBE, Infectiology
Institute, University of Muenster and Ivan Liashkovich
Institute for Physiology II, University of Muenster
❚ Leica AF7000 – The premium solution for highly demanding applications which offers full real-time control for fast 3D time-lapse
experiments, TIRF, Fura2, FRET SE, deconvolution and peripheral
triggering.
EGF stimulated endocytosis
Green: GFP tagged EGF-receptor
Courtesy of: Vibor Laketa, Cell Biology and Biophysics
Unit, EMBL Heidelberg
5-day-old transgenic zebrafish larva
Green: GFP – actin
Courtesy of: Prof. Dr. Stephan C. F. Neuhauss,
Institute for Brain Research, ETH Zurich
3
A System as Dynamic
as Modern Research
The new Leica Advanced Widefield Systems offer full flexibility
to adapt to your changing needs. No matter which modules and
components you select to configure an individual system, you will
benefit from the utmost reliability and convenience. As many of
the system components are designed and developed by Leica
Microsystems, we offer unparalleled automation control due to
complete integration of our hardware within the software platform.
The basic Leica AF6000 E system can be upgraded with a software
package to create a Leica AF6000 fluorescence and imaging analysis system. Add a control unit to convert the system into a highend Leica AF7000 live cell workstation with full real-time control.
“I need to perform high-resolution
3D-deconvolution microscopy
in time while keeping the cells in
optimal conditions. Leica Microsystems provides the right tool
for my live cell research.”
Due to the many different fluorescence applications, all systems
from the Leica AF6000 upward work exclusively with semi- or fully
automated microscopes. Leica Microsystems technical experts
perform the upgrade on site at your facility, which minimizes the
inconvenience of system down-time.
Dr. Eef Parthoens
Department for Molecular Biomedical
Research (DMBR), VIB - Ghent University,
Ghent, Belgium
>>
4
>>>
Upgrade Options from Entry-Level to High-End Solutions
Leica AF6000 E
Leica AF6000
Leica AF6500
Leica AF7000
Main Characteristics Entry-level fluorescence
acquisition system,
2D time-lapse, multichannel overlay
Fluorescence imaging
and analysis system,
high flexibility (without
real-time control)
High-speed fluorescence Dedicated live cell system,
system, real-time control full real-time control
of camera and one filter
wheel
Applications
Fluorescence imaging
and documentation
General fluorescence
imaging, 3D time-lapse
experiments, multi-positioning, deconvolution
Functionality of AF6000,
plus fast 2D time-lapse
and fast ratio imaging,
peripheral triggering
TIRF, fast 3D time-lapse,
Fura2, FRET, deconvolution, peripheral triggering
Includes
AF6000 E SW
+ PC
AF6000 SW
+ PC
AF6000 SW
+ PC + CTR6500 HS
AF6000 SW
+ PC + CTR7000 HS
Leica DM1000
Ergonomic system
microscope
Leica DM2000
Advanced system
microscope
Leica DM2500
Advanced system
microscope, 100 W
Leica DM3000
Advanced system
microscope, motorized
Leica DM4000 B
Automated upright
microscope for biological
laboratory and research
applications
Leica DM5000 B
Automated upright
microscope for demanding
research applications
Leica DM5500 B
Automated upright
microscope for high-end
research applications with
motorized focus
Leica DM6000 B
Fully automated upright
microscope for cutting
edge biomedical research
Leica DMI3000 B
Manual inverted research
microscope
Leica DMI4000 B
Automated inverted
research microscope
Leica DMI6000 B
Fully automated inverted
research microscope
Leica M205 FA
High-end stereomicroscope
for fluorescence imaging
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5
Leica AF6000 E –
Flexibility Comes First
|||
Entry-level image acquisition system
|||
Document your research
|||
Combine with a wide range of
Leica cameras and microscopes
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2D time-lapse and
multi-channel overlay
The Leica AF6000 E is the ideal entry-level solution for widefield
fluorescence microscopy. It is particularly suited to general fluorescence imaging applications such as biological and cytomorphological examinations and for professional documentation of
research results.
The system is truly flexible, as it can be teamed with both manual
and automated Leica Microsystems microscopes. In addition, the
Leica AF6000 E can be configured with the Leica M205 FA, Leica
Microsystems’ automated stereo fluorescence microscope. The
system is compatible with many cameras in the Leica Microsystems range, which benefit from full integration with the software.
As with all systems in the Leica Advanced Widefield System Line,
the Leica AF6000 E features an intuitive user interface and intelligent workflow guidance.
Mouse kidney section
Glomerulus
Blue: DNA, DAPI
Green: Alexa 488 WGA
Red: Actin, Alexa 568 Phalloidin
Image by:
Dr. M. Beynon, Leica Microsystems, Milton Keynes, UK
Leica AF6000 E with DM2500 upright microscope and Leica EL6000 external light source
6
Leica AF6000 –
Truly Flexible for Fluorescence
The Leica AF6000 provides all the necessary facilities for demanding fluorescence applications. This flexible, fully integrated system
can be equipped with a large number of application packages and
software modules.
The Leica AF6000 application spectrum ranges from general fluorescence work via multi-channel imaging to time-lapse experiments,
multi-positioning and z-stacking. The system incorporates a wealth
of standard features for image documentation, processing, quantification and analysis.
|||
Multi-dimensional 3D imaging
|||
Live cell experiments
|||
A range of application packages
and software modules for further
analysis
Optional modules for FRET, advanced time-lapse, channel unmixing, colocalization and various deconvolution techniques will
extend the analysis capability.
COS1 cell culture
Red: Galectin3-YFP
Green: Caveolin1-CFP
Courtesy of: Prof. Dr. R. Jacob, University of Marburg,
Department of Clinical Cytobiology and Cytopathology,
Marburg, Germany
Leica AF6000 with DM6000 B upright microscope and climate chamber
7
Leica AF6000 – for Upright,
Inverted and Stereo Microscopes
Flexibility for a wide range of applications
The Leica AF6000 is compatible with automated upright and inverted microscopes from Leica Microsystems. In addition, the Leica
M205 FA fluorescence stereomicroscope with FusionOptics™ is
a perfect partner for the Leica AF6000, with applications in timelapse and 3D image capture in the world of developmental biology.
The use of a motorized stage also enables documentation of images
on several selected areas of interest in the specimens.
Vascular anatomy of a zebrafish embryo as revealed
by GFP expression driven by the Fli-1 promoter.
The Leica AF6000 can be combined with monochrome and color
cameras as well as EMCCD cameras. The camera of choice for
high-resolution fluorescence imaging is the Leica DFC360 FX.
Courtesy of: Dr. Brant Weinstein,
National Institutes of Health, Bethesda, MD
Leica AF6000 with M205 FA stereomicroscope and Leica EL6000 external light source
8
Leica AF6500 –
Ultrafast 2D Time-lapse
The Leica AF6500 is a high-speed fluorescence system that allows
real-time control of a camera and a filter wheel as required for
fast 2D time-lapse studies. Automated research microscopes,
both upright and inverted, from Leica Microsystems are the ideal
platforms for a Leica AF6500.
Use the ratio imaging function to measure concentrations of second messenger substances such as Ca2+ at resolution over long
periods of time. The system is also ideal for examinations of a
wide variety of cellular processes such as exocytosis, transport
processes within the cytoskeleton and membrane fusions.
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Real-time 2D time-lapse experiments
|||
Multi-dimensional 3D imaging
|||
High-speed ratio imaging
|||
Peripheral triggering
C. elegans
GFP expression
Image by:
Dr. Werner Wittke, Leica Microsystems, Wetzlar, Germany
Leica AF6500 with DM6000 B upright microscope, EMCCD camera and Leica EL6000 external light source
9
Leica AF7000 – Real-time 3D
Time-lapse Applications
|||
Real-time 3D time-lapse experiments
|||
High-speed FRET SE experiments
|||
TIRF experiments
The Leica AF7000 leaves nothing to be desired in widefield fluorescence imaging. This system combines the ultimate in speed,
reliability, scope for experiment design, analysis facilities and operational convenience. Achieve excellent results – fast!
This extremely versatile and fully integrated system allows realtime control of all necessary hardware components for complex
live cell studies, such as the camera, fine focus devices and up to
four external filter wheels. In addition, it offers the possibility to
observe live cells using TIRF. With the Leica AF7000 you are expertly equipped for the research tasks of today and tomorrow.
The Leica AF7000 effortlessly performs fast 3D time-lapse experiments. It is also fully capable of performing FRET SE, TIRF, as well
as fast Fura2 measurements. Trigger signals can be transmitted to
peripheral components via four channels. Vice versa, trigger signals
can be received in up to four channels from external components
and used to start experiments.
Co-migration of the zebrafish mechanosensory lateral
line placode (green) and innervating axons (red) at 36
hours post fertilization. Live embryo is a double transgenic carrying neuronal-specific and placode-specific
constructs (NBT:dsRedExpress and claudinB:mGFP,
respectively). z-stack 200 slices/80 μm. Processing
steps: dye separation and deconvolution.
Courtesy of: Dr. Darren Gilmour, EMBL,
Heidelberg, Germany
Leica AF7000 with DMI6000 B inverted microscope, climate chamber, CO2 controller and SuperZ Galvo focus
10
TIRF (Total Internal Reflection Fluorescence) is the ideal technique
for the study of cellular membranes and their environments. With
automatic laser adjustment to maintain constant TIRF penetration
depth, the Leica AM TIRF MC offers maximum flexibility for TIRF
applications with multiple fluorophores at wavelengths of 405 nm,
488 nm, 561 nm and 635 nm for multi-color applications. The individual laser lines are coupled and controlled quickly and precisely
via AOTF (Acousto-Optical Tunable Filters), achieving a switching
time of only 1 ms. The Leica AF7000 is the base workstation from
which to upgrade to the Leica AM TIRF MC at a later date.
COS1 cell culture, Widefield
Red: Galectin3-YFP
Green: Clathrin-CFP
COS1 cell culture, TIRF
Red: Galectin3-YFP
Green: Clathrin-CFP
Courtesy of: Prof. Dr. R. Jacob, University of Marburg,
Department of Clinical Cytobiology and Cytopathology, Marburg, Germany
“Cell Biology is a fascinating field
of research with a wide range of
applications. My equipment needs
to grow with the increasing number of demanding applications –
Microsystems
just like the Leica Microsystem
imaging solutions.”
Prof. Dr. R. Jacob
University of Marburg, Department of C
Clinical
Cytobiology and Cytopathology,
Marburg, Germany
Leica AM TIRF MC – visualize life‘s secrets with true multi-color TIRF
11
Design Your Experiment –
with Intelligent Modules
Mitosis observed
DIC
H2A-YFP
No mitosis observed
DIC
H2A-YFP
t1
Common to all Leica Advanced Widefield Systems, Leica Microsystems has developed an easy-to-use, intuitive software interface that guides you through every step of the work process. Incorporating the latest findings in ergonomic design and efficiency
research, the software enables you to define experiments and
analysis swiftly and effortlessly.
t2
t3
t4
t5
t6
From the simple control of the microscope functions to the highly
complex confocal systems, all the fluorescence research systems
are built on the same software platform. This saves time when you
first start to work, getting your results fast.
All settings are stored so experiments can easily be repeated. Log
files and image parameters give detailed information on all actions
performed during an experiment.
A context-sensitive help function is available whenever needed.
Support for complex analysis procedures comes in the form of
specific wizards. For example, FRET SE offers application-oriented
help and explanations.
||| Mark
t7
NIH 3T3 mouse fibroblast cells expressing H2A-YFP.
Two positions of a Mark and Find experiment.
Courtesy of: Dr. Constantin Kappel, DKFZ, Heidelberg, Germany
and Find
– Activation of the live image during the experiment
– Change the cell positions while the experiment is in progress
||| Live
Data Mode
– Change the course of an experiment “on the fly”
– Online generation and graphical display of measurements
– Flexible definition and combination of various
experimental routines
– Peripheral in/out triggering of experiments;
control of manipulators or peristaltic pumps
||| 3D
Visualization
– 3D rendering and dynamic user interaction of volume
– Compile multi-channel 3D movies
Colocalization module: 2D-cytofluorogram
12
||| Colocalization
– Visualization and quantification of colocalized
fluorochromes
– Statistic determination of the degree of colocalization
||| Dye
Finder
– Elimination of cross-talk between fluorochromes
with a proprietary unmixing algorithm
– Differentiation between dyes with overlapping spectra
||| Deconvolution
– Removal of haze and blur by reassigning scattered light
– Fully integrated deconvolution algorithm using adaptive,
calculated or supplied PSF
Widefield image stack
Deconvolved image stack
Cerebellum (nervous system), fixed sample, 40 μm thickness
Courtesy of: Dr. Fulvio Florenzano, S. Lucia Foundation, Rome
||| FRET
–
–
–
–
SE
Application module for the acquisition and analysis
of FRET specimens
Correction of cross-talk
Online display of FRET image and FRET graph
FRET Sensitized Emission and FRET ratio
||| Fura2
– Determination of Ca2+ concentrations for intracellular signal
processing and transport processes
– Ca2+ quantification according to Grynkiewicz formula
– Online generation of measurement curves and cleaning
of background fluorescence
||| Well
Plate Acquisition
– Predefined patterns for 6, 12, 24 and 96 microplates
– User-definable patterns
– Combination with time-lapse, z-stacking and tile scan
Fura2 software module: increasing Ca++ concentration in
the cell by adding Bradykinin
Ratio intensity chart of Fura2 time-lapse experiment
13
Optimum Conditions
for Live Cells
Live cell experiments need optimized hardware components. Key
requirements include fast or real-time image recording, temperature stability, minimum exposure of specimens to light and highly
sensitive recording techniques for weak fluorescence. For successful studies and high-quality results, all parameters have to be optimized for the specimens being analyzed and the experimental
conditions.
||| Wide choice of cameras from Leica Microsystems, supplemented
Leica Microsystems offers a wide choice of hardware components
and accessories for individual configuration of your research system. The modules can be combined in many different ways. The
full integration of all automated functions with the software ensures perfect interaction of components in everyday use.
||| Wide choice of microscope stages ranging from manual through
by highly sensitive EMCCD cameras. The Leica DFC360 FX is the
premier high-resolution, high-performance fluorescence camera
recommended for the majority of Leica AF7000 applications. The
Leica DFC360 FX offers overlapping mode for fast real-time studies,
excellent signal-to-noise ratio, 40 MHz readout rate for real-time
exposures and a highly sensitive CCD.
motorized 3-plate stages to scanning stages.
Extensive range of accessories for live cell examinations including climate chamber, cell chamber, various manipulators, CO2
controller, incubators.
|||
||| Fast external filter wheels and filter sets from Leica Microsystems
with extremely short switching times.
||| Leica
EL6000 external light source with high-speed shutter for
minimum light exposure.
||| Leica
TIRF objective for high spatial and temporal resolution in
studies of dynamic processes inside or close to the cell membrane.
||| SuperZ
focus for ultra-fast and highly precise z-stacking.
High-Speed Components for the Leica Advanced Widefield System Line
Time-lapse
Multi-channel overlay
Camera control
Leica AF6000 E
●
●
●
Z stack
Motorized XY stage
Deconvolution
External filter wheels
● 1 FW*
Leica AF6000
●
●
●
●
●
●
● 1 FW*
Peripheral triggering
SuperZ Galvo fine focus
Pifoc fine focus
TIRF
●● available, high-speed ● available, non high-speed
14
not available
Leica AF6500
●●
●●
●●
●
●
●
●● 1 FW
●● 2 In, 1 Out
Leica AF7000
●●
●●
●●
●●
●
●
●● 4 FW
●● 4 In, 4 Out
●●
●●
●●
*Leica CTR5500/6000/6500 required
Leica External Fast Filter Wheel for high-speed excitation,
attenuation and emission control.
Leica STP6000 Smart Touch Panel for complete and
convenient control of the microscope.
Leica DFC360 FX high-resolution camera for fast fluorescence imaging.
Leica EL6000 external light source with high-speed shutter
for fluorescence excitation.
Leica HCX Plan Apo objective with 1.46 numerical aperture for TIRF applications.
SuperZ Galvo focus for ultra-fast and highly precise z-stacking.
15
Leica Microsystems –
the brand for outstanding products
• Life Science Research Division
Leica Microsystems’ Life Science Research Division supports the imaging needs of the scientific community with
advanced innovation and technical expertise for the visualization, measurement and analysis of microstructures.
Our strong focus on understanding scientific applications
puts Leica Microsystems’ customers at the leading edge of
science.
• Industry Division
The Leica Microsystems Industry Division’s focus is to
support customers’ pursuit of the highest quality end
result by providing the best and most innovative imaging
systems for their needs to see, measure and analyze the
microstructures in routine and research industrial applications, in materials science and quality control, in forensic science investigations, and educational applications.
• Biosystems Division
The Biosystems Division of Leica Microsystems brings
histopathology labs and researchers the highest-quality,
most comprehensive product range. From patient to
pathologist, the range includes the ideal product for each
histology step and high-productivity workflow solutions for
the entire lab. With complete histology systems featuring
innovative automation and Novocastra™ reagents, the
Biosystems Division creates better patient care through
rapid turnaround, diagnostic confidence and close customer collaboration.
• Surgical Division
The Leica Microsystems Surgical Division’s focus is to
partner with and support micro-surgeons and their care
of patients with the highest-quality, most innovative surgical microscope technology today and into the future.
Leica Microsystems’ mission is to be the world’s first-choice provider of innovative solutions to our
customers’ needs for vision, measurement and analysis of micro-structures.
Leica, the leading brand for microscopes and scientific instruments, developed from five brand
names, all with a long tradition: Wild, Leitz, Reichert, Jung and Cambridge Instruments. Yet Leica
symbolizes innovation as well as tradition.
Leica Microsystems – an international company
with a strong network of customer services
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