Leica DM6000 CFS User Manual

Leica DM6000 CFS

Confocal Fixed Stage System

The solution for applications in live cell research, from physiology
to electrophysiology – from single cells to whole organisms

Leica TCS SP5 with integrated fixed stage micro­scope Leica DM6000 CFS

Excellent Results
Under All Conditions

Leica Microsystems sets a new standard with the integration of the
Leica DM6000 CFS fixed stage microscope into the Leica TCS SP5
confocal platform.

Neurobiological research has a long history not only in measure­ments of single cells, but also of thick brain slices. Here more
realistic results can be achieved due to the intact network. Ulti­mately, accessing the brain directly in a whole animal provides
the most intact environment for conducting measurements on
individual cells. So for optimal results, space for whole animal
samples is an absolute must.

Electrophysiological research employs micropipette systems for
recording electrical signals (patch clamp, whole cell recordings),
electrical stimulation (intracellular stimulation, synaptic stimulation,
soma-stimulation), dye injection and intracellular perfusion. To make
such measurements effective, the microscope needs to have as
much space as possible for micropipettes and other manipulators.

Features and Benefits

Maximum workspace for mani -

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pulators and attached pipettes

High collection efficiency due to

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short collection path before

detectors

Integrated change of magnification

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(0.35x, 1x, 4x) of camera port

Remote control of all microscope

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functions via touch panel

Special booster optic to fill the

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entrance pupil of the 20x1.0

objective

“Dip-in” feature for immersing the

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objective

Patented condenser drainage

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system

High microscope and sample

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stability

Searching for the perfect spot in the sample requires a large field
of view, whilst the precise positioning of the micropipettes requires
high magnification. Furthermore, high quality confocal image scan­ning requires a large numerical aperture for best performance. To
fulfill these experimental prerequisites, a camera with an
adjustable magnification and specially adapted objectives are
needed. Any direct manipulation of the system potentially disturbs
the delicate positioning of the micropipettes within the sample.
This emphasizes the importance of a remote control for the imag­ing setup, providing convenient access to all relevant functions.

Finally, recording electrophysiological and imaging data with per­fect synchrony is paramount for correct interpretation. Triggering
image recording by external events and synchronizing the appli­cation of stimuli with image scanning down to the single line lev­el helps realize sophisticated experimental setups. Online analy­sis of the resulting data allows on-the-fly adjustment of external
parameters, helping the researcher to get everything just right.

The Leica TCS SP5 with the integrated fixed stage microscope
Leica DM6000 CFS gives excellent experimental results under all
conditions.

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Variable Field
at High Resolution

With the highest aperture objective 20x 1.0 NA, with the innova­tive, ergonomic and stable microscope stand Leica DM6000 CFS,
with short coupled non-descanned detectors and with the fastest,
most sensitive scanning system SP5, we enter a new dimension
in imaging, from single cells to whole organisms.

The objective

The new Leica HCX APO L 20x 1.0 water immersion objective
offers both a large field and a high resolution with one single
objective lens, making it unnecessary to change objectives
between overview imaging for sample preparation and detail
imaging for data recording.

In addition to optimizing the optical parameters, a number of
physical, chemical and mechanical improvements have been
integrated into the new HCX APO L 20x1.0 application objective: It
is corrosion-free, chemically neutral and avoids diffusion of met­al ions. Wettability is outstanding, thermal conductivity is minimal
and magnetic fields at the front of the objective have been elimi­nated.

Leica objective HCX APO L 20x1.0

Transmission HCX Apo L 20x/1.00 W

These new objective characteristics are possible by employing a
special, extremely hard ceramic material for the whole front lens
area. This material is resistant to mechanical damage and much
better suited to the requirements of electrophysiology than typical
objective metal sleeves.

A further advantage is the access angle of the objective, which is
a measure of how easily manipulators can be fitted. This has been
widened to 39 degrees, which is almost the limit of what is theo­retically possible. In combination with a free working distance of
2 mm, the 20x objective is ideal for many applications.

Features and Benefits

20x water immersion objective

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High numerical aperture (NA) of 1.0

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Free working distance (FWD) of

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2 mm

Optimized access angle of 39°

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Large objective focusing range of

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13 mm

Insulated ceramic tip

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Minimum conductivity

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High transmission in VIS and IR

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(see graph)

Best DIC and Dodt contrast

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Leica DFC350 FX Camera with parfocal magnifi­cation changer

The microscope stand

But an objective alone doesn’t make an electrophysiology setup.
The Leica DM6000 CFS combines the specific needs of electro­physiological experiments with the optimized imaging perfor­mance of the DM series. With its focusing objective or nosepiece,
it offers a highly stable and convenient optical platform for all
fixed stage applications.

To cover the demand for large scale sample screening and fine
needle approaches for electrophysiology, the DM6000 CFS has a
special observation tube with 3 magnification positions (0.35x; 1x;
4x). This parfocal magnification changer allows a total change in
magnification up to a factor of 12 without changing the objective.

The specimen is completely decoupled from the microscope, both
mechanically and electrically. The microscope provides a high
degree of free space for application-specific sample holders,
bath chambers and several manipulators at a time. In this way,
the sample and the patch electrodes can be moved below the
fixed optical axis of the microscope. This allows the researcher to
scan the entire dendritic tree and follow the axons of fluores­cently labeled cells. The DM6000 CFS is also designed for use
with a separate third-party fixed platform for holding micromanip­ulators and other devices, as well as an x,y-translation stage.

Features and Benefits

of the Leica STP6000

Intuitive user interface

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Control of all microscope functions

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Z-focusing wheel for coarse and

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fine z-movements

Freely programmable function keys

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Helps to avoid vibrations caused by

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manual operation

To avoid the need for direct manipulation of the microscope stand
which could cause accidental vibrations, the external control panel
Leica STP6000 (Smart Touch Panel) controls all the microscope
functions. A touch screen and freely programmable function keys
allow quick and easy operation without adversely affecting the
measurements.

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