Microscopy from Carl Zeiss

LSM 5 Family
goes ZEN

Efficient Navigation in
Laser Scanning Microscopy

We make it visible.
LSM 5 Family

The fifth generation of Laser Scanning Microscopes

from Carl Zeiss offers efficient solutions for every
confocal application.

The family comprises a wide range of models to

match every need, be it single-user instruments or
multi-user research facilities, single color or multi
color spectral imaging, UV/Visible imaging wave-
lengths or multiphoton microscopy, slow to ultrafast
imaging or fluorescence correlation spectroscopy.

The members of the LSM 5 Family consist of 4 build-

ing blocks. The Microscope, the Laser Module, the
scanning and detection unit and the new LSM
Software: ZEN. Discover the ideal combination of
precision and intuition, and experience the beginning
of a new era in Laser Scanning Microscopy.
System Concept 2

ZEN Software 4

LSM 5 EXCITER 6

LSM 510 8

LSM 510 META 10

LSM 510 NLO 12

ConfoCor 3 14

LSM 5 LIVE 16

LSM 5 DUO 18

Techniques and Methods 20

Functions and Tools 22

Figure Legends 23

Brochures 25

Microscope :
The body
The Laser Scanning Microscopes of the LSM 5 Family
can be configured around different Carl Zeiss
research microscopes to suit different applications.
All motorized functions of these high end research
microscopes are software controlled. Their integra-
tion into a uniform overall optical concept war-
rants optimum performance and prime imaging
quality – simply: optical perfection.
2
+ +
Laser module :
The source
Depending on the application, the Laser Scanning
Microscopes of the LSM 5 Family are equipped
with a variety of lasers – from UV through the
visible range to the near infrared. With the con-
venient software control, lasers can be activated
and attenuated with the speed and precision
needed to minimize photodamage, which is most
important for highly sensitive specimen.

A Perfect Kit of Building Blocks
+ +
Scanning and detection module :
The heart
Independently movable scanning mirrors in the
scanning and detection modules exactly direct the
laser beam across the specimen, permitting free
adaptation of the scanfield to the specimen
requirements in terms of shape, size and orientation.
Depending on model and application, the scanning
and detection module may have up to four photo-
multiplier detectors. For the requirements of multi-
fluorescence microscopy, in particular, Carl Zeiss
designed the META detector. This is a multichannel
detector used for recording the spectral emission
profile of every spot of the specimen (“Spectral
Imaging”). In conjunction with intelligent software
algorithms, the META detector exactly separates
combinations of fluorochromes having extremely
similar emission properties.
System Concept
New software – ZEN:
The brain
The Laser Scanning Microscopes are controlled by the
LSM Software. For the LSM 5 Family, a total innovation
in user friendly software is available: ZEN. Implemen-
ting new programming and design tools, every soft-
ware feature has been redeveloped to be more intuitive
to use. It enables an operator to rearrange the user
interface exactly as required and save all user preferences.
The sophisticated functionality of ZEN is supplemented
by optional software packages tailored to specific
applications. Additional licenses for workstations are
of course available.
3
 Intuition is the Key
With intuitive usability, entirely new
possibilities for customization and
many other innovative features,
ZEN – Efficient Navigation, the new
software for the LSM 5 Family enables
users to focus exclusively on what is
relevant for their work: the specimen
to analyze. The software provides users
with an intelligible, clearly organized
interface at the same time as it accom-
modates the highest demands of
scientific research.
Off-line software
Image Browser
The Image Browser is an optional software
package available free of charge. Download it from
web site www.zeiss.de/lsm.
Use the browser for viewing, editing, sorting,
printing and exporting LSM 5 images.
Additional licenses
An additional license includes the full functionality
of the LSM 5 Software and allows you to set up
another workstation.
Optional software packages
3D for LSM
• Visualization of 3D images
• Analyze/Measure 3D images
Deconvolution
• Image restoration from 3D image data
• Ability to compute Point Spread Functions
• Visualization of the computed image data
4
Multiple Time Series
• Acquire and visualise complex dynamic processes
• Automatic image acquisition in multiple positions
• Image acquisition with different configurations for every
ROI, also in combination with bleaching
Image VisArt plus
• Fast 3D/4D reconstruction and animation
• Shadow, Transparency and Surface rendering techniques
• Presentation and animation of the images
• 3D Measurements
Visual Macro Editor
• Programing of complex experiments with the help of
graphical elements
• Using the Visual Macro Editor you can use image data in a
feedback loop to control image acquisition
Physiology
• Synchronize acquisition and local bleaching e.g. for FRAP
• Mean ROI analyses and tracks intensity changes over time
• Calibration and measurement of ion concentrations
FRET
• Reproducible acquisition and analysis of FRET experiments
• Sensitised Emission and Acceptor photobleaching techniques
• Calculations include Youvan, Gordon & Xia formula
 ZEN Software

The Central Screen

Area serves as the
stage for imaging.
Here too, the clear
and intuitive layout
makes the interface
user friendly. You
can edit display
options for multi-
dimensional data
View Control Panels are located below the sets using the View-
image. These include all the tools needed to Buttons on the left
interact with a data set. The visibility of side of each image.
the View Control Panels can be toggled to
simplify the interface during an experiment.

The Z-Stack Tool enables you to easily

navigate through an image stack and
simultaneously control your microscope.
Both features have been visually
designed so that they are completely
intuitive to use.

Benefits

Straightforward system control and

configuration via the graphic user interface
Individual adaptation to the experiment
by intuitive user-defined configuration
Reproducibility with a mouse click –
the ReUse function quickly restores all image
acquisition parameters
Clearly structured data visualization and analysis
with easy-to-handle tools
New and intuitive software design
Individual adaptation of the software interface
for quick restoring of different software layouts.

5
 Entry-Level Confocal Microscopy

The LSM 5 EXCITER is a powerful confocal system for demanding

routine tasks in biomedical laboratories, education and research.
Full motorization, superb image quality, multitracking, and
comfortable software all combine to make it unique in its class.
The extraordinary variability of the LSM 5 EXCITER gives you
unprecedented choice of application.

Configuration Applications

Microscopes 3D analyses of cell and tissue structures

Upright: Axio Imager.Z1, Axio Imager.M1, Multiple fluorescence and quantitative
Axioskop 2 FS MOT
colocalization experiments, e.g. neurogenetic
Inverted: Axio Observer.Z1
studies (Fig. 7.1)
+ Laser modules
Wavelength ranges:
Monitoring of fast physiological processes by
Fast Ion Imaging, e.g. when measuring calcium
405 nm – 543 nm or 458 nm – 633 nm

+ Scanning modules
LSM 5 EXCITER scanning module with one or two
detection channels, an adjustable pinhole + AOTF.
Options: Transmitted-light channel
transients (Fig. 7.2)
Identification and quantification of molecule
interactions by FRET microscopy
Analysis of protein motility with FRAP

+ Software
Basic software ZEN for LSM 5 EXCITER
Options: Physiology, Image VisArt plus, Decon-
volution, 3D for LSM, Macro recorder/editor, FRET,
FRAP, Visual Macro Editor, Multiple Time Series

6
 LSM 5 EXCITER

7.1

7.2

Benefits

Exact repetition of instrument imaging conditions with

the ReUse software function
Sequential Imaging of spectrally overlapping fluorescence
signals (crosstalk) by the frame or line
multitracking concept
Observation of fast biological processes
thanks to high scanning speeds and flexible
image formats
Finest image quality due to adjustable and
variable pinhole
Pro/Basic-Mode can be selected for each tool indivi-
dually with the press of a button.

7
 That Extra Bit of Individuality

The LSM 510 is ideal for solving complex analytical

problems in cross-disciplinary research teams,
multi-user facilities, microscopy service centers,
or in the central analytical lab. Wherever you need
that extra bit of system performance, flexibility or
add-on modularity. With its excitation wavelengths
ranging from 351nm to 633nm, fast, pixel-
accurate scanfield definition, a proven multiple
pinhole concept and many more features,
the LSM 510 is bound to convince you.

Configuration Applications

Microscopes Efficient analysis of complex multiple fluores-

Upright: Axio Imager.Z1, Axio Imager.M1, cence signals with a broad spectrum of
Axioskop 2 FS MOT fluorochromes, e.g., the structural relationship
Inverted: Axio Observer.Z1 between cell compartments in organs (Fig. 9.2)

+ Laser modules
Wavelength ranges: 351/405 nm – 633 nm
Direct observation of transport processes in live
cells and organisms by targeted photoactivation

+ Scanning modules
LSM 510 scanning module with up to four detection
channels and up to four adjustable pinholes
Option: Transmitted-light channel and additional
coupling of manipulator LSM DuoScan
(for photoactivation, photoconversion and uncaging).
of novel fluorochromes such as Kaede (Fig. 9.1)
Study of diffusion and transport mechanisms by
targeted photobleaching experiments
Determining the molecular interactions of
biomolecules by sensitized emission or
acceptor bleaching in FRET microscopy

+ Software
Basic software for LSM 510
Options: Physiology, Image VisArt plus, Decon-
volution, 3D for LSM, Macro recorder/editor, FRET,
FRAP, Visual Macro Editor, Multiple Time Series

8
 LSM 510

9.1 9.2

Benefits

Fast acquisition of image stacks of multiply

labeled specimens in up to four detection channels
Selective bleaching and imaging of a specimen
segment with the RealROI concept
Precise results in multiple fluorescence
experiments due to identical detection volumes
High flexibility in using fluorochromes thanks to
a broad spectrum of excitation wavelengths
Workspace zoom helps to bridge the distance between
microscope and monitor.

9
 Spectral Dimensions

Add another dimension: The LSM 510 META recognizes fluorescence

signals by their unique spectral signatures. The Emission Fingerprinting
technique provided by Carl Zeiss positively and precisely separates
spectrally overlapping fluorescence signals because the system analyzes
the entire spectrum of the fluorochrome.

Configuration Applications

Microscopes Exact separation of fluorochromes with

Upright: Axio Imager.Z1, Axio Imager.M1, extremely overlapping emission spectra, such as
Axioskop 2 FS MOT Texas Red and Cy3 (Fig. 11.2)
Inverted: Axio Observer.Z1 Gene expression studies and protein
+ Laser modules
Wavelength ranges:
localization in biological systems, especially by
Spectral Imaging of multiple fluorescent proteins
351/405 nm – 633 nm or 458 nm – 633 nm Spectral imaging of autofluorescence, as in
+ Scanning modules
LSM 510 META scanning module with two
corals (Fig. 11.1)
Examination of protein-protein interactions
conventional and 32 spectral detection channels by FRET microscopy in conjunction with Emission
and three individual pinholes Fingerprinting
Option: Transmitted-light channel and additional
Observation of dynamic processes and imaging
coupling of manipulator LSM DuoScan
of ion concentrations by measurement of the
(for photoactivation, photoconversion and uncaging).
spectral changes of fluorescence signals
+ Software
Basic software for LSM 510.
Precise analyses of multiple fluorescence
in situ hybridization (M-FISH)
Options: Physiology, Image VisArt plus, Decon-
volution, 3D for LSM, Macro recorder/editor, FRET,
FRAP, Visual Macro Editor, Multiple Time Series

10
 LSM 510 META

11.1 11.2
Relative Intensity

Wavelength (nm)

Benefits

Time-saving, specimen-preserving spectral

imaging by parallel data acquisition in multiple META
channels
Unambiguous, reliable separation of
fluorescent signals by the proprietary Emission
Fingerprinting technique
Detection of the spectral properties
of fluorochrome and autofluorescence signals
On-line separation of fluorescence during measure-
ments by the Online Fingerprinting technique
Multiple viewing options can easily be chosen in the
versatile Central Screen Area.

11
 Deep Insights

The LSM 510 NLO is ideal for the sensitive

in-depth analysis of live specimens, including
whole organisms. The LSM 510 NLO is out-
standing for its unparalleled selectivity:
Even low concentrations of fluorochromes can
be detected due to precisely tunable multi-
photon excitation and non-descanned detection.
With the depth-selective excitation unique to
the LSM 510 NLO, your bleaching experiments
will succeed with pinpoint accuracy.

Configuration Applications

Microscopes 3D and 4D microscopy of thick tissue sections

Upright: Axio Imager.Z1, Axio Imager.M1, and complete organisms, e.g. in the study of
Axioskop 2 FS MOT. Inverted: Axio Observer.Z1 structures in a zebra fish or in the cortex of
+ Laser modules
Wavelength range: Visible: 458 nm to 633 nm
a mouse (Figs. 13.1, 13.2)
Live cell microscopy of specimens with high
IR: 690 to 1064 nm (depending on laser) photosensitivity, as of a live zebra fish (Fig. 13.3)
+ Scanning modules
LSM 510 scanning module with one to three
Highly 3D-localized uncaging of biologically
active molecules from caged compounds
detection channels
Z-selective bleaching experiments thanks to
Option: LSM 510 META scanning module with
the 3D effect of multiphoton excitation
two conventional and 32 spectral detection channels
and three individual pinholes Separation of fluorescence signals by their fluores-
Options: Up to two reflected light NDD detectors and cence lifetimes: Fluorescence Lifetime Imaging
up to two transmitted light NDD detectors or coupling Microscopy (FLIM)
of manipulator LSM DuoScan
(for photoactivation, photoconversion and uncaging)

+ Software
Basic software for LSM 510
Options: Physiology, Image VisArt plus, Decon-
volution, 3D for LSM, Macro recorder/editor, FRET,
FRAP, Visual Macro Editor, Multiple Time Series

12
 LSM 510 NLO

13.1 13.2 13.3

Benefits

Highly informative imaging of fluorescence

signals in tissue layers hundreds of microns deep
by femtosecond multiphoton excitation
Reduced phototoxicity thanks to near infrared
excitation
Efficient detection of weak fluorescence
signals by non-descanned detection (NDD) in
reflection and transmission
Correct for scattering with depth by automatic
adaptation of laser power to specimen penetration depth
Flexible use of various fluorochrome combinations
by elimination of excitation crosstalk using Excitation
Fingerprinting
Intuitive interaction with
the microscope through the graphic
representation of the Z-Stack.

13
 Analysis of Moving and
Blinking Molecules

The ConfoCor 3 in combination with the LSM 510 META represents a

fully integrated imaging and spectroscopic platform for single-molecule
detection. The system analyses complex processes with high statistical
significance. Either in solution or in a cell, the ConfoCor 3 will provide
reliable information about concentration, diffusion constants or binding
kinetics.

Configuration Applications

Microscope Analysis and localisation of molecular inter-

Inverted: Axio Observer.Z1 actions within a cell

+ Objectives
C-Apochromat 40 x W NA 1.2
Measurement and imaging of extremely low
molecule concentrations
C-Apochromat 63 x W NA 1.2 In vivo studies of molecular mobility to
LD C-Apochromat 40 x W NA 1.1 analyse cellular diffusion and transport processes
+ Scanning modules
Imaging: LSM 510 with up to three conventional
Investigation of association and dissociation
processes for the functional characterization
channels or LSM 510 META with up to two of cellular proteins
conventional and 32 spectral detection channels
Determination of binding kinetics in biochemistry
Spectroscopy: ConfoCor 3 with two detection
and biophysics
channels for photon counting imaging

+ Detectors
LSM 510: PMT ConfoCor 3: APD

+ Software
LSM 510: Basic software plus options including
Physiology, MultipleTime Series, Image VisArt plus, Decon-
volution, 3D for LSM, Kinetics, FRET, Visual Macro Editor
ConfoCor 3: Basic software plus options including
Extended Models, Global and Interactive Fit, and
Photon Counting Histogram

14
 ConfoCor 3

15.2

15.1

EGFP-CENP-A

15.3

Benefits

Integrated platform for confocal imaging and

spectroscopic analysis
Maximum sensitivity and time resolution
based on high performance detectors
Real-time analysis based on intelligent
algorithms
High information density by assessing
localization, concentration, interaction and diffusion
of molecules within one measurement
Pre-defined models for fast and flexible data evalution.

15
 Visions of Life

The LSM 5 LIVE is a revolutionary parallel scanning

confocal design that offers new levels of speed
and sensitivity to capture life in motion like never
before.
The line scanning approach allows image dimensions
and parameters to be perfectly balanced for every
sample. Imaging calcium transients at hundreds
of XY frames per second for example or imaging fine
structural changes at several Z-stacks per second,
the LSM 5 LIVE is extremely versatile and efficient.

Configuration Applications

Microscopes Capture fast moving structures such as blood

Upright: Axio Imager.Z1, cells or versicles with fast 4 dimensional image
Axioskop 2 FS MOT acquisition
Inverted: Axio Observer.Z1 Confocal high speed imaging in 4 Dimensional
+ Laser modules
Wavelength ranges: 405 – 635 nm
Biology even with low magnification lenses
Perform physiology measurements perfectly
+ Scanning modules
LSM 5 LIVE scanning module with one or two high
matched to the biological timescales
Visualise fine structures at high resolution with
sensitivity line detectors the necessary sensitivity
Option: Additional coupling of manipulator unit
Study subcellular dynamics by pixel-precise
LSM DuoScan (for photoactivation, photoconversion
bleaching, photoactivation, photoconversion and
and uncaging)
uncaging with the LSM 5 LIVE DuoScan
+ Software
Basic software for LSM 5 LIVE.
Options: Physiology, Image VisArt plus, 3D for LSM,
Multiple Time Series, Macro recorder/editor, FRET,
FRAP, Visual Macro Editor

16
 LSM 5 LIVE

17.1 17.2

Intensity ROI 1

Time [s]

Benefits

Ultrafast acquisition – Observe dynamic

processes in the kilohertz range
Imaging precision – Optical images with
outstanding 3-dimensional resolution
Unmatched sensitivity – thanks to a ground-breaking
beam splitter, the AchroGate, which achieves over 95 %
efficiency of both excitation and emission
Effortless data handling – of Gigabytes of data with
real time electronics and computing
Easy set up and control of the system due to the
clear visual representation of the Light Path.

17
 The Ultimate Confocal Workstation

The LSM 5 DUO provides an optimal solution for

multiuser facilities and extremely demanding samples.
By combining all current top range confocal technologies,
the user gets fast line scanning, ultraprecise point
scanning, spectral and non-linear imaging plus flexible
sample manipulation all together in one system for the
first time. Controlled by the same software and
realtime electronics, sharing
powerful lasers and the
microscope, this is a
highly efficient setup.

Configuration Applications

Microscopes Developmental studies with high speed and

Upright: Axio Imager.Z1, confocallity even with low magnification lenses
Axioskop 2 FS MOT
Perform physiology measurements both
Inverted: Axio Observer.Z1
perfectly matched to the biological timescales
+ Laser modules
Wavelength ranges:
or the spectral properties
Exact separation of fluorochromes with
351 – 635 nm or 405 – 1050 nm

+ Scanning modules
LSM 5 LIVE, LSM 510, and LSM 510 META
extremely overlapping emission spectra, including
autofluorescence signals

scanning modules with three to six dedicated Sample manipulation experiments like
confocal detectors FRAP, FLIP, FLAP, Photoactivation, Photoconversion

+ Software
Basic software for LSM 5
Options: Physiology, Image VisArt plus, 3D for
LSM, DCV, Multiple Time Series, Macro recorder/
editor, FRET, FRAP, Visual Macro Editor
and Uncaging with precision and high resolution
timescales
Capture fast moving structures such as blood
cells or versicals with fast 4 dimensional image
acquisition

18
 LSM 5 DUO

19.1 19.2
A t=0 s B t=7 s

Intensity ROI 1
C t = 10 s D t = 30 s

Time [s]

Benefits

Ultrafast acquisition – Observe dynamic

processes in the kilohertz range
Imaging precision – Optical images with
outstanding 3-dimensional resolution
Time-saving, specimen preserving spectral imaging –
by parallel data acquisition in multiple META channels
Sample manipulation – two independent
scanner groups allow great flexibility for
photobleaching, activation and conversion
Unmatched sensitivity – thanks to a ground-
breaking beam splitter, the AchroGate, which achieves
over 95% efficiency of both excitation and emission
Effortless data handling – of Gigabytes of
data with real time electronics and computing
User-friendly multi-dimensional imaging
enabled by clearly arranged view options and
functional view control panels.

19
 LSM 5 EXCITER

LSM 510 META

LSM 510 NLO

Techniques and

LSM 5 DUO
LSM 5 LIVE
ConfoCor 3
LSM 510
Methods

3D Investigations
Added information in three spatial dimensions (X,Y,Z) by using
the confocal volume:
Acquisition, visualization and measurement of 3D image stacks. ■ ■ ■ ■ ■ ■ ■

FRET by Sensitized Emission (Fluorescence Resonance Energy Transfer)

Investigation of molecule interactions by energy transfer between
fluorescence-labeled donor and acceptor molecules spaced at 1–10 nm:
Direct registration of FRET by detecting acceptor fluorescence intensity ■ ■ ■ ■ ■ ■ ■
after donor excitation.

Ion Imaging
Measurement and imaging of ion concentrations by means of selective
fluorescence markers:
Image acquisition, measurement and calibration of ion concentrations. ■ ■ ■ ■ ■ ■ ■

Multi-fluorescence Imaging
Image generation from specimens with multiple fluorescence labels:
Crosstalk-free detection and presentation of multiple fluorophores.
■ ■ ■ ■ ■ ■ ■

Quantitative Colocalization
Detection of the coincidence of two fluorescence-labeled molecules
in the confocal detection volume. Investigation of neighborhood
relations and interactions: Definition of parameters, image presentation ■ ■ ■ ■ ■ ■ ■
and data analysis (colocalization coefficients).

Time Series
Added information on simple dynamic processes by acquisition
of image series, also in combination with local bleaching:
Acquisition, visualization and analysis of time series (X,Y,t or X,Y,Z,t). ■ ■ ■ ■ ■ ■ ■

FLIP and FRAP Microscopy (Fluorescence Loss in Photobleaching;

Fluorescence Recovery after Photobleaching)
Investigation of diffusion and transport mechanisms by targeted
bleaching under observation: ■ ■ ■ ■ ■ (■1) ■
Image acquisition and analysis of the bleaching experiments.

FRET with Acceptor Bleaching (Fluorescence Resonance Energy Transfer)

Investigation of molecule interactions by energy transfer between
fluorescence-marked donor and acceptor molecules spaced at 1–10 nm:
Indirect registration by detection of acceptor and donor fluorescence ■ ■ ■ ■ ■ (■1) ■
intensities after acceptor bleaching.

20
 LSM 5 EXCITER

LSM 510 META

LSM 510 NLO

LSM 5 DUO
LSM 5 LIVE
ConfoCor 3
LSM 510
Complex Time Series
Acquire additional information about complex dynamic processes by simultaneous
automated acquisition of image series in several freely defined ROIs, also in ■ ■ ■ ■ ■ (■1) ■
combination with local bleaching:
Acquisition (at variable intervals, in different positions, with different scanning
modes and configurations), visualization and analysis of complex time series.

Uncaging, Photoactivation, Photoconversion

Activation or uncaging of biologically active substances by intense irradiation:
Targeted, pixel-accurate irradiation in a region defined in spatial and temporal ■ ■ ■ ■ ■ (■1) ■
terms, followed by imaging of the sample.

Emission Fingerprinting & Spectral Imaging

Acquisition of spatially resolved fluorescence emission spectra.
Acquisition of lambda stacks for emission fingerprinting, presentation and saving ■ (■2) (■3) (■4)
of fluorescence spectra.
Online processing and presentation of spectrally unmixed, multichannel images.

Spectral Unmixing Clear separation of potentially heavily overlapping

fluorescence and autofluorescence signals using a variety of techniques:
Emission Fingerprinting & Reference Spectra, (■3) (■3) ■ (■2) (■2) (■3) (■4)
Emission Fingerprinting & User-Defined Spectra of ROIs,
Emission Fingerprinting & Automatic Component Extraction,
Automatic Channel Unmixing.
Optical Manipulation
Optional coupling of additional scan head (LSM DuoScan) to enable
imaging parallel to photoactivation, photoconversion and uncaging ■ ■ ■ ■ (■5)
within freely defined ROIs.

FLIM Microscopy (Fluorescence Lifetime Imaging Microscopy)

Measurement of the fluorescence lifetime properties of fluorochromes.
Spatially resolved measurement of fluorescence intensity as a function ■ (■4)
of time.

Multiphoton Microscopy
Fluorescence excitation through multiphoton processes.
For the penetration of thick specimens and specimen-sparing long-time ■ (■4)
experiments: Excitation with femtosecond pulsed infrared laser,
image acquisition, and data analysis.

FCS (Fluorescence Correlation Spectroscopy)

Statistical analysis method for the quantitative study of molecule interactions,
based on the fluctuation of fluorescence-marked molecules in the confocal volume: ■
Data acquisition and analysis.

1 2
= Bleach ROI restricted to Rectangle. = If scanhead has the META detector option.
3
= Only Channel based unmixing available – not suitable for the separation of very closely overlapping spectra.
4
= Functionality of the LSM 5 DUO Systems varies depending on the constituent systems. 5
= Functionality of all LSM 5 DUO Systems. 21
 Functions and Tools

ZEN Software Functionality for all Systems of the LSM 5 Family

Auto Z
On-line adaptation of acquisition parameters for Z stacks to equalize brightness in the various Z sections.
Pro-Basic-Concept
Reduced complexity of all tools – full functionality upon a mouse-click.
Channel Mode
Sequential excitation of the specimen to obtain multiple fluorescence images without emission crosstalk.
ReUse
Restitution of acquisition parameters by a mouse click, for reproducible experiments.
Tile Scan
Acquisition of a mosaic consisting of partial images, for recording larger objects.
Z Bleach
Bleaching in a defined specimen plane for targeted manipulation.

ZEN Software Functionality for all Point Scanning Systems of the LSM 5 Family (except the LSM 5 LIVE)

Crop
Convenient graphical selection of scanning regions; zoom and rotation for the recording of fast processes.
Spline Scan
Scanning along a freely defined line for capturing fast processes.
Spot Bleach
Defined bleaching in a specimen spot, for observation of extremely fast dynamic movements.
Spot Scan
Scanning with extremely high temporal resolution of signal intensity in a confocal spot.
Real ROI Scan
Acquisition and imaging of freely defined specimen regions, with laser blanking for minimum light load on the specimen.
ROI Bleach
Defined bleaching in freely defined specimen regions, for applications such as FRAP, FRET, photoactivation,
photoconversion, uncaging.
Step Scan
Fast overview scanning, with skipped rows replaced by interpolation.

Specification of Systems of the LSM 5 Family

Point Scanners: LSM 5 EXCITER/LSM 510/

Maximum scanning resolution
Maximum scanning speed
Example of High Frame Rate
Scanning rotation
Data depth
Line Scanner: LSM 5 LIVE
LSM 510 META/ LSM 510 NLO
2048 x 2048 pixels 512 x 2048 pixels
5 frames/s at 512x512 pixels, or 2600 lines/s 120 frames/s at 512x512 pixels, or 30,000 lines/s
77 frames/s at 512x32 pixels 1010 frames/s at 512x50 pixels
Free 360° rotation in steps of 1° None
Choice of 8 or 12 bits

22
Images in this brochure

7.1) Atrium of a guinea pig. Actin
(BODIPY FL phallacidin: green) and
NeuroPeptide-Y (Alexa 594: red).
Specimen: Prof. Y. Satoh,
Iwate Medical University, Japan
7.2) Salivary gland of the fly Calliphora vicina.
Hormone-induced increase in cytosolic calcium
concentration, imaged with Fluo-4 in the intact
organ.
Specimen: Dr. B. Zimmermann,
Potsdam University, Germany
15.1) HEp-2 cells expressing the centromeric
protein CENP-A fused to EGFP.
FCS measurements were taken at the indicated
position in the nucleoplasm. Bar indicates 5 µm.
15.2) Record of the intensity fluctuations.
15.3) Autocorrelation function and fit of the data
to a one component anomalous diffusion model
reveal the transport coefficient.
Specimen: Peter Hemmerich,
Leibnitz Institute for Age Research – Fritz Lipmann
Institute, Jena, Germany

9.1) Photoconversion of Kaede transfected living

17.1) Zebrafish embryo. Erythrocytes (dsRed : Red)
cell by repeated local irradiation at 405 nm.
and endothelial cells (eGFP : Green)
Specimen: Prof. A. Miyawaki, Riken, Japan
2 Channels captured simultaneously at
9.2) Morphology of a plant root. Epidermis (blue), 33 frames / second.
mitochondria (red) and cell walls (green). Specimen: Dr. S. Hermanson and Dr. S. C. Ekke,
Orthogonal sections of a Z stack. University of Minnesota, USA
Specimen: Prof. P. C. Cheng,
17.2) Rat, ventricular cardio myocytes (Fluo-4,
New York State University, Buffalo, USA
Ca2+ ion indicator) & mean ROI plot Captured
at 300 frames / second.
Specimen: Dr. J. Lederer,
11.1) Autofluorescent pigments of the coral
Medical Biotechnology Center, Baltimore, USA
Lobophyllia corymbosa. Spectral signatures in the
indicated ROIs show GFP-type coral proteins and
chloroplasts of dinoflagellate symbionts.
19.1) Time course of the intracellular distribution
Specimen: Dr. A. Salih, Australian Key Centre for
of photoconverted Kaede after single irradiation
Microscopy & Microanalysis,
with 405 nm in ROI 1.
University of Sidney, Australia
(A) Before conversion, the whole cell is green-
11.2) HeLa cells with Trans Golgi network (GCC-
fluorescent.
GFP: blue), microtubules (Cy3: green), actin (Texas
(B) After selective irradiation at 405 nm of ROI 1
Red: red), visualized by Emission Fingerprinting.
inside the nucleus, the converted Kaede very quickly
Specimen: Prof. J. Stow and J. Lock,
spreads throughout the nucleus (C and D).
University of Queensland, Australia
19.2) FRAP experiment with ROI bleaching and
fast linescan of a GFP labelled cell.
13.1) Zebra fish embryo. Optical section through
head and eye, double labeled with antibodies
against axonal cell surface components.
Specimen: Prof. M. Bastmeyer, Jena University,
Germany
13.2) Mouse cortical neurons with dendrites
and spines expressing YFP. Maximum projection
of 50 images.
Specimen: M. Fuhrmann and J. Herms,
Neuropathology, LMU München, Germany
13.3) Spinal cord of a living transgenic zebra fish
embryo expressing GFP in motoneurons and their
processes.
Specimen: Dr. M. Marx, Prof. M. Bastmeyer,
Jena University, Germany

23
Patents

LSM 5 EXCITER
US Patents: 5127730, 6037583, 6167173, 6462345,
6563632, 6665068, 6848825
German Patents: 19702752C2, 69131176T2

LSM 510
US Patents: 6462345, 6486458, 6167173, 6278555, 6037583,
6563632, 6631226, 5127730, 6848825, 6941247
German Patents: 19758744C2, 19758745C2, 19758748C2, 19702753C2,
19702754C2, 19702752C2, 19827140C2, 69131176T2
EP Patent: 0977069B1

LSM 510 META

US Patents: 6403332, 6750036, 6858852, 6747737, 7009699
German Patents: 19915137C2, 10038526B4

LSM 510 NLO

US Patents: 6178041, 6521899, 5034613, 6344653, 6403332,
6867915, 7119898
German Patents: 19919091C2, 69032621T3, 69034117T2

ConfoCor 3
US Patents: 6591223, 6693742
EP Patent: 1183525B1

LSM 5 LIVE
US Patents: 6848825, 6888148, 6947127
EP Patent: 1617264B1

LSM 5 DUO
US Patent: 7212337

24
Ask for the following LSM 5 EXCITER
brochures which give Laser Scanning Microscope
detailed information on
each system of the LSM 510 and
LSM 5 Family. LSM 510 META
For further imformation, Laser Scanning Microscopes
please visit us at
www.zeiss.de/lsm
LSM 510 NLO and
LSM 510 META NLO
Multiphoton Laser Scanning Microscopes

ConfoCor 3
Detection Module for Fluorescence Fluctuation Analysis

LSM 5 LIVE and

LSM 5 LIVE DuoScan
Laser Scanning Microscopes

LSM 5 DUO and

LSM DuoScan
Laser Scanning Microscope Workstations

Mikroskopie von Carl Zeiss Mikroskopie von Carl Zeiss

LSM 5 EXCITER LSM 510 Mk4 und LSM 510 METAMk4

Laser Scanning Mikroskop Laser Scanning Mikroskope

Die erste Wahl

Zelluläre Prozesse verfolgen
für Ihre zellbiologische Forschung

We make it visible. We make it visible.

45-0070 45-0066 45-0009

Mikroskopie von Carl Zeiss

LSM 5 LIVE und LSM 5 LIVE DuoScan

Laser Scanning Mikroskop

Visionen kommen in Bewegung

We make it visible.

45-0057 45-0067 45-0056

25
Perfection is not an Art,
it is an Attitude

The precision and performance of our devices are a direct

result of our pursuit of technological perfection.
Our devices have paved the way for many significant
discoveries and scientific breakthroughs.
Microscope systems made by Carl Zeiss continue to provide
the basis for a better understanding of the process of life
and the world around us.

For further information, please contact:

Printed on environment-friendly paper,
bleached without the use of chlorine.

Carl Zeiss MicroImaging GmbH

07740 Jena, Germany
Phone: +49 3641 64 3400
Subject to change.

Fax: +49 3641 64 3144

E-mail: micro@zeiss.de

www.zeiss.de/lsm 45-0073 e/08.07