Leica TCS SP5 X User Manual

Leica TCS SP5 X

X-citing Life – The Only Confocal That Tunes Into Any Excitation

Ready for any dye the future brings

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The only system that fully adapts to the sample

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Full freedom and flexibility in emission AND excitation

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All you need for imaging – today and tomorrow!

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Biological and medical research fluorescence
imaging has evolved from studying one or two
features in a specimen to examining even more
features and their dynamic interactions in the
cellular environment. This requires a variety of
fluorescent dyes to study four, five, or more pro­teins simultaneously. As new dyes are devel­oped and used, the ability to optimally excite
and detect these fluorophores is necessary in
both living and fixed specimen.

Until now, research in multi-user facilities or
institutes has been limited by the type or num­ber of dyes that could be excited. Conventional
laser scanning imaging systems are equipped
with a set of gas or solid state lasers that emit
fixed laser lines for use. Using this equipment
the researcher has to choose the dyes and
labels for detection according to the imaging
system. The experiment has to be compromized
to the available instrumentation.

Leica TCS SP5

X-citing Life!

The Leica TCS SP5 X supercontinuum confocal
unites the broadband capabilities of the Leica
TCS SP5 AOBS®and the freedom and flexibility
to select any excitation line within the continu­ous range of 470 to 670 nm.

With our patented spectral detection technolo­gy you have complete freedom to choose the
detection area in up to five confocal PMT chan­nels. The acousto-optical beam splitter AOBS
is the only option for a supercontinuum laser,
and allows the selection of up to eight simulta­neous lines from anywhere in the white light
spectrum.

®

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“The White Light Laser

The Only System that
Fully Adapts to the Sample

Why should researchers be limited because the dyes for imaging
can not be excited sufficiently? Why should limitations be placed
on the laser configuration choices for a multi-user core facility?

The Leica TCS SP5 X features a fully tunable supercontinuum
laser that provides excitation lines from 470 nm to 670 nm in the
visible spectrum. Easy control of excitation lines is at your finger­tips. Freedom to choose the specific excitation line in 1 nm incre­ments allows optimal adjustment of the excitation to the special
properties of the dye. This flexibility decreases damage to the cell
and reduces unwanted cross-excitation.

offers a tremendous
flexibility and will open
our minds to new imaging
applications.”

Dr. Maria C. Montoya

Confocal Microscopy and Cytometry Unit
Biotechnology Program
Spanish National Cancer Center (CNIO)
Madrid, Spain

The Leica TCS SP5 X is the only confocal system that adapts to the
research needs of today and tomorrow.

Leica Design by Christophe Apothéloz

visible and ultraviolet radiation:

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Intuitive Interface Allows Full Freedom
and Flexibility for Scanning

• Excitation lines can easily be selected to any wavelength with 1 nm accuracy

• Tuning via the LCD control panel allows to select excitation light while scanning

• Easy intuitive interface makes the Leica TCS SP5 X perfect for basic and advanced users

• Interface allows easy access to all scanning parameters for both supercontinuum and fixed laser lines

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AOBS®allows any combination of

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8 laser lines – perfect for super-

continuum laser

Greater sensitivity than any

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other confocal

AOTF-controlled laser power

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Combination of fixed laser lines and

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supercontinuum lines together

poses no problem

Opening Doors and Opening M

Ready for any new dye of the future

The number of available fluorescent dyes is growing tremen­dously. The ability to excite these dyes optimally with ideal spec­tral detection and minimal sample damage from high excitation
sources is challenging. For most imaging applications the num­ber of dye pairs available is endless, however, the ability to prop­erly excite and control cross-excitation is a difficult procedure
with the limited fixed laser lines available in today’s laser scan­ning confocal systems.

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100 –

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“The supercontinuum
laser and AOBS are a
perfect match. Full
freedom of a white light
laser could only be
realized with the AOBS.
The Leica TCS SP5 X
saves experimental time
to focus more on
data analysis.”

Prof. Alberto Diaspro

Department of Physics
University of Genoa
Genoa, Italy

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40 –

20 –

0

The TCS SP5 X perfectly adapts to any dye that will come today or
tomorrow. With 200 nm of freely tunable excitation lines, the Leica
TCS SP5 X can be tuned to provide optimal excitation of any avail­able dye. This flexibility is critical for core facilities that service hun­dreds of users with various samples and various dyes. There will
never be a need to turn away any sample or have difficult times
exciting new dyes.

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A new dimension to spectral separation

New GFP mutants are all perfectly separated by excitation map­ping with the Leica TCS SP5 X

Cy2, exc 491 nm, emis 499 – 505 nm,
NMDA receptors

Lp, exc 474 nm, 509 – 514 nm,
beads in vacuols

Acridine Orange, exc 484 nm,
emis 520 – 532 nm, nucleus

Cy3, exc 551 nm, emis 555 – 575 nm,
Gaba-B-R1 receptors

TexasRed, exc 562 nm, emis 605 – 612 nm,
vacuoles and endosomes

Alexa 594, exc 594, emis 615 – 625 nm,
nuclear and cellular membrane

Up to 5 spectral detection channels

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Tune to exact spectral range while

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scanning

Almost no light loss from first to last

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channel

Cooled PMTs allow for greater

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sensitivity and less noise

Never worry about barrier filters

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again

Best red emission detection

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available

Emission and excitation scans allow

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for new dimensions in spectral

imaging

2 years of scanning light guaranteed

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Alexa 633, exc 633 nm, emis 655-665 nm,
clathrine vesicles

Cy5, exc 640 nm,
emis 661-675 nm, cilia

Paramecium stained with 8 fluorescent dyes. Excitation and emission lambda
scans were obtained to determine optimal imaging conditions for each dye.
Courtesy of P. Ramoino and P. Bianchini, LAMBS-MicroScoBio, Dept. of Physics,
University of Genoa, Italy

Most confocal systems on the market claim to have some capabil­ity to “spectrally” separate dyes. The Leica TCS SP5 X allows the
use of emission scan dye separation and excitation scan dye sep­aration for a new dimension in spectral imaging.

This technology opens new doors to confocal microscopy appli­cations including excitation AND emission mapping that can be
used for new dye development, or existing applications such as
FRET.

“With the TCS SP5 X we
are solving problems we
did not know existed.”

Dr. Kees Jalink

Department of Cell Biology H1
The Netherlands Cancer Institute
Amsterdam, The Netherlands

What can the Leica TCS SP5 X do for you?

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Leica Microsystems –
the brand for outstanding products

Leica Microsystems operates internationally in four divi­sions, where we rank with the market leaders.

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