Zeiss VivaTome User manual

VivaTome
Operating Manual

Carl Zeiss COPYRIGHT VivaTome
2 423639-9000-701 M60-2-0046 e 05/2012
Knowledge of this manual is required for the proper operation of the instrument. Would you therefore
please make yourself familiar with the contents of this manual and pay special attention to instructions
for safe handling of the device.
Subject to change due to technical progress; the manual is not covered by an update service.
© Unless expressly authorized, forwarding and duplication of this document, as well as utilization and
communication of its contents are not permitted. Violations will entail an obligation to pay
compensation.
All rights reserved in the event of granting of patents or registration of a utility model.
Issued by: Carl Zeiss Microscopy GmbH
Carl-Zeiss-Promenade 10
07745 Jena, Germany
microscop[email protected]m
www.zeiss.com/microscopy
Carl Zeiss Microscopy GmbH
Königsallee 9-21
37081 Göttingen, Germany
Manual number: M60-2-0046 e
Date of issue: Version 6, 01.05.2012

VivaTome CONTENTS Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 3
CONTENTS
Page
1Notes on Instrument Safety ............................................................................................ 4
1.1 General Indications............................................................................................................. 4
1.2 Safety Notes....................................................................................................................... 5
1.3 Warning and Information Labels on the Device.................................................................... 8
1.4 Warranty Notes .................................................................................................................. 8
2DESCRIPTION.................................................................................................................... 9
2.1 Intended Use...................................................................................................................... 9
2.2 Functional Principle............................................................................................................. 9
2.3 Description of the Instrument and Main Features............................................................... 11
2.4 System Overview VivaTome............................................................................................... 12
2.5 Technical Data.................................................................................................................. 13
3START-UP ....................................................................................................................... 15
3.1 Installation Requirements.................................................................................................. 15
3.2 Requirements as to the Microscope Stand ......................................................................... 16
3.3 Basic Microscope Settings ................................................................................................. 17
3.4 Mounting VivaTome ......................................................................................................... 17
3.5 Mounting Supporting Feet................................................................................................ 18
3.6 Mounting the Camera ...................................................................................................... 18
3.7 Connecting VivaTome....................................................................................................... 19
4OPERATION .................................................................................................................... 20
4.1 Operational and Functional Elements ................................................................................ 20
4.2 Switching On ................................................................................................................... 20
4.3 Controlling the Scanhead via AxioVision............................................................................ 21
4.4 Changing the Filter Module .............................................................................................. 21
5CARE, MAINTENANCE, TROUBLESHOOTING AND SERVICE ......................................... 23
5.1 Care of the Device............................................................................................................ 23
5.2 Maintaining the Device ..................................................................................................... 23
5.3 Troubleshooting ............................................................................................................... 24
5.4 Service Request ................................................................................................................ 24

NOTES ON INSTRUMENT SAFETY
Carl Zeiss General Indications VivaTome
4 423639-9000-701 M60-2-0046 e 05/2012
1NOTES ON INSTRUMENT SAFETY
1.1 General Indications
VivaTome has been designed, manufactured and tested in compliance with the standard DIN EN 61010-1
(IEC 61010-1) "Safety Regulations for Electrical Measuring, Control and Laboratory Instruments".
The instrument meets the requirements of the EC Directives 2006/95/EC "Low-Voltage Directive" and
2004/108/EC "Electromagnetic Compatibility", as well as the standard DIN EN 62471 (IEC 62471)
"Photobiological Safety of Lamps and Lamp Systems".
It carries the mark.
The instrument has to be disposed of in compliance with the WEEE Directive 2002/96/EC.
The warning and information symbols explained below will help you recognize at a glance the type and
purpose of the notes included in this operating manual:
CAUTION
This symbol indicates a possible hazard to the user of the instrument.
CAUTION
Do not look into the light source! This could cause damage to your eyes. Risk class 2 according
to IEC 62471
CAUTION: High-energy UV radiation!
Risk of damage to eyes and skin!
CAUTION
Disconnect the power cable before opening the instrument!
ATTENTION
This symbol indicates a possible hazard to the instrument or system.
NOTE
This symbol indicates a note containing special instructions.
Apart from the present operating manual you should observe the following operating manuals for your
detailed information:
−Operating manual of VivaTome manufactured by Aurox Ltd.
−Operating manual of the microscope (Axio Imager, Axio Observer or Axio Examiner)
−Operating manual for the computer system
−Operating manual of the camera (AxioCam MRm, HRm or EM)
−Operating manual of HXP 120 C or HXP 120

NOTES ON INSTRUMENT SAFETY
VivaTome Safety Notes Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 5
1.2 Safety Notes
VivaTome including its original accessories must not be used for applications other than those described
in the present operating manual.
Please observe the following warnings:
Follow the instructions given in the manufacturers' operating manuals of the HXP
120 and
VivaTome!
Observe the operating manual of the microscope in use to ensure reliable functioning of the
illumination system in combination with the microscope.
Use the instrument for Carl Zeiss microscopes only.
The manufacturer cannot assume any liability for other applications, including those of
individual modules or single components. This also appl
ies to any service or repair work that is
not carried out by authorized service personnel. In case of non-compliance, all warranty claims
shall be forfeited.
Do not operate the device and the accessories included in the delivery in potentially explosive
areas, in the presence of volatile anesthetics or flammable solvents, such as alcohol, benzine or
similar chemicals.
The VivaTome power supply unit is classified as a protection class II unit (with protective
insulation) and must not get in contac
t with moisture. If its casing is damaged, put the power
supply unit out of operation.
The instrument is not equipped with special devices for the protection from substances that are
corrosive, potentially infectious, toxic or radioactive, or from
other substances that could be
hazardous to health. Make sure to observe all legal requirements, particularly the relevant
national accident prevention regulations when handling such substances.
If equipped with a white light source coupled by a liq
uid light guide, the instrument emits
ultraviolet radiation that may cause burns to the eyes and skin. Therefore, never look directly
into the light and avoid direct incidence of the light on your skin. When operating the
microscope, always use the protective devices belonging to the instrument.
Never look into the light beam - neither with nor without optical instruments, even not if you
simply want to observe the specimen. Your eyes may be damaged in case of non-observance!

NOTES ON INSTRUMENT SAFETY
Carl Zeiss Safety Notes VivaTome
6 423639-9000-701 M60-2-0046 e 05/2012
According to the int
ended use, only a 100/0 deflection to the camera port with mounted
VivaTome is provided.
Never look directly into the light guide output of the white light source to avoid the risk of
being dazzled and going blind.
The white light source must not be switched on until after having connected the output of the
liquid light guide tightly with VivaTome and VivaTome firmly screwed to the coupling point of
the microscope.
Take appropriate protective measures if the specimen releases noxious gases, dust, and vapors,
secondary radiation or explosive substances as a result of the UV radiation!
When using the white light source HXP 120 / HXP 120 C, please pay attention to the
manufacturer's operating manual.
The bulb of the external white light source has to be changed according to the manufacturer's
operating instructions of the white light source. Otherwise there is danger of burns and
explosion when changing the bulb.
Maintenance or repair work, as well as any modification, removal or exchange of components
not described in this operating manual, or any other intervention in the equipment may only be
carried out by Carl Zeiss Microscopy GmbH staff or by persons
expressly authorized by the
company.
Please take into consideration that VivaTome is a high-precision opto-
electronic system.
Improper handling can easily impair its functioning or even damage it and will lead to a forfeit
of all warranty claims.
Use VivaTome in closed rooms only.
Dirt and dust may impair the performance of the devices. They must therefore be protected
from such influences to the greatest possible extent.
In order to prevent vibrations during the operation, an anti-vibration plate or a vibration-
damped system table should be used in connection with VivaTome.
Read in any case the safety data sheet for Immersol 518 F®.

NOTES ON INSTRUMENT SAFETY
VivaTome Safety Notes Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 7
The immersion oil Immersol 518 F®
irritates the skin. Avoid any contact with skin, eyes and
clothing.
After skin contact, wash the oil off with plenty of water and soap. After eye contact,
immediately rinse the eye with plenty of water for at least five minutes. If the irritation persists,
consult a medical specialist.
Proper disposal of Immersol 518 F®
: Take care to ensure that it does not enter surface water or
the sewage system.
Do not touch the hot lamp housings and adhere to a cooling time of about 15 minutes.
Defective devices must not be disposed of with the household waste; dispose of
them in
compliance with the provisions of the law.
Also specimens have to be disposed of properly according to the applicable legal provisions and
internal work instructions.

NOTES ON INSTRUMENT SAFETY
Carl Zeiss Warning and Information Labels on the Device VivaTome
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1.3 Warning and Information Labels on the Device
Pay attention to the warning and information labels attached to VivaTome and the microscope. Please
check the device for completeness of all labels shown below and contact the Carl Zeiss Microscopy
GmbH if one of the labels is missing. You will receive a substitute free of charge.
Fig. 1-1 Warning and information labels on VivaTome
1.4 Warranty Notes
The manufacturer guarantees that the device is free from material or manufacturing defects when
delivered. Possible defects must be notified immediately and steps be taken to minimize damage. If
notified of such a defect, the manufacturer is obligated to rectify it at its discretion, either by repairing
the instrument or by delivering an intact replacement. No guarantee is provided for defects caused by
natural wear (wearing parts in particular) and improper use.
The instrument manufacturer shall not be liable for damage caused by faulty operation, negligence or
any other tampering with the microscope, particularly the removal or replacement of device components,
or the use of accessories from other manufacturers. This shall lead to a forfeit of all warranty claims.
No maintenance or repair work other than the operations mentioned in the present manual may be
carried out to VivaTome. Repairs are only permitted to the Carl Zeiss Service staff or to persons
specifically authorized by this department. In case of fault please turn first to the Carl Zeiss representative
competent for you.

DESCRIPTION
VivaTome Intended Use Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 9
2DESCRIPTION
2.1 Intended Use
Product name: VivaTome
VivaTome is a system for creating and imaging optical sections in real time with the aid of a so-called
spinning disk provided with a grid pattern. As light source, an HXP 120 / HXP 120 C is used.
The images are captured with a CCD camera in real time.
Use the scanhead only in combination with the microscope stands Axio Observer, Axio Imager or
Axio Examiner.
2.2 Functional Principle
VivaTome creates optical sections according to the "aperture correlation" technology that was first
presented by T. Wilson and collaborators at Oxford University. This technology combines the speed of a
spinning disk approach with the light efficiency of structured illumination.
The core of VivaTome is a spinning disk in a conjugate image plane, through which the illumination light
passes to the specimen and the emission light from the specimen is detected. To obtain optical sections,
the disk is structured in a precisely defined way, providing the excitation light with a structure that is
imaged sharply only in the focus. The emission light from the specimen is detected again through the
same disk.
In VivaTome, highest light efficiency is of utmost importance. The structure of the VivaTome spinning disk
allows up to 50 % of the incident excitation light to reach the specimen, so that a simple white light
source is sufficient. This is why the emission detected through the disk is not immediately an optical
section, but the information from the focus plane is superimposed with a conventional widefield image.
To calculate an optical section from this image, a second image, containing exclusively the widefield
contribution, is needed. For this purpose, the disk surface facing the specimen is mirrored. Light reflected
by the disk comes from areas above and below the focus plane, constituting exactly the information
required to calculate the optical section. This light is also imaged onto the camera through a split-view
optics, so that a single shot contains all the information needed to calculate an optical section.
To ensure optimal light efficiency, the disk does not contain individual pinholes, but a grid pattern
structure. Two different grid patterns are available to choose from in order to achieve optimal results with
different objectives. Both grid patterns are arranged optimally on the disk, excluding transition artifacts
between the sectors.

DESCRIPTION
Carl Zeiss Functional Principle VivaTome
10 423639-9000-701 M60-2-0046 e 05/2012
Calculation based on partial images:
The image detected through the disk contains mainly intensities from the focal plane, however
superimposed with a widefield portion. In order to obtain an optical section, this widefield intensity must
be subtracted. For this purpose, the system uses the second, simultaneously detected image, which was
reflected from the disk and thus contains "out-of-focus" intensities. An optical section is obtained by
simple scaled subtraction of both images:
imagetionedconfocalalconventionKconfocalalconvention sec)()( =−×−+
In this case, Krepresents a fixed constant correcting differences in the light efficiency of both imaging
pathways.
A scaled addition of both images yields a conventional widefield image. Thus, all the information from
the specimen is captured by VivaTome in a raw data image and is available for further analysis.

DESCRIPTION
VivaTome Description of the Instrument and Main Features Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 11
2.3 Description of the Instrument and Main Features
The following components are required for operating VivaTome:
−Microscope Axio Observer, Axio Imager or Axio Examiner
−Camera adapter 60N-C 1" 1.0x
−High-resolution and high-aperture objectives, e.g. Plan-Apochromat 63x/1.4 Oil
−PC with at least 2 GB working memory
−Monitor for image display
−Digital camera AxioCam MRm, AxioCam HRm, AxioCam EM
−Image editing software AxioVision (minimum Rel. 4.8)
−Vibration-damped table or anti-vibration plate (Axio Observer and Axio Imager)
−HXP 120 or HXP 120 C illumination
VivaTome is mounted to the side port (Axio Observer) or camera port (Axio Imager and Axio Examiner) of
the microscope using a c-mount camera adapter.
Four different filter sets are available.
Light is delivered through a liquid light guide.
1Filter module
2VivaTome
3Camera adapter matching the filter module
Fig. 2-1 VivaTome with filter module and camera adapter

DESCRIPTION
Carl Zeiss System Overview VivaTome VivaTome
12 423639-9000-701 M60-2-0046 e 05/2012
2.4 System Overview VivaTome
Fig. 2-2 System overview

DESCRIPTION
VivaTome Technical Data Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 13
2.5 Technical Data
Dimensions of VivaTome (W x D x H)................................................approx. 280 mm x 150 mm x 70 mm
Dimensions of power supply unit (W x D x H) .....................................approx. 120 mm x 55 mm x 40 mm
Dimensions of HXP 120 / HXP 120 C (W x D x H)............................approx. 130 mm x 210 mm x 290 mm
Weight of VivaTome ......................................................................................................................6.4 kg
Weight of power supply unit..........................................................................................................0.2 kg
Ambient conditions
Transportation and storage (in packaging)
Admissible ambient temperature........................................................................................ -10 to +50 °C
Admissible air humidity (no condensation) ............................................................20 % to 75 % at 35 °C
Atmospheric pressure ....................................................................................................800 to 1060 hPa
Operation
Admissible ambient temperature....................................................................................... +10 to +35 °C
Admissible relative humidity................................................................... 20 % to 80 %, no condensation
Atmospheric pressure ..............................................................................................800 hPa to 1060 hPa
Pollution degree ................................................................................................................................... 2
Operating data
VivaTome
Supply voltage....................................................................................................... 12 V DC; max. 1.25 A
Power supply unit VivaTome
Line voltage........................................................................................................100 to 240 VAC ±10 %
Change of line voltage unnecessary!
Line frequency...........................................................................................................................50/60 Hz
Power consumption of power supply unit.................................................................................... 400 mA
Output voltage ......................................................................................................... 12 VDC; max. 1.3 A
Environment for use ............................................................................................................closed rooms
Protection class......................................................................................................................................II
Protection type .................................................................................................................................IP20
Electrical safety........................................................................................in compliance with EN 61010-1

DESCRIPTION
Carl Zeiss Technical Data VivaTome
14 423639-9000-701 M60-2-0046 e 05/2012
HXP 120/HXP 120 C
Line voltage ....................................................................................................... 100 to 240 VAC ±10 %
Change of line voltage unnecessary!
Line frequency...........................................................................................................................50/60 Hz
Maximum power consumption ............................................................................................max. 210 VA
Fuses ...................................................................................................................................2x T 2.5 A/H
Environment for use ............................................................................................................closed rooms
Protection class ......................................................................................................................................I
Protection type................................................................................................................................ IP 20
Electrical safety............................................................in compliance with DIN EN 61010-1 (IEC 61010-1)
Optical Data
Light output VivaTome - microscope................................................. risk group 2 according to IEC 62471
Light guide coupling VivaTome ........................... risk group 2 according to IEC 62471 (HXP 120 coupling)

START-UP
VivaTome Installation Requirements Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 15
3START-UP
Read in any case the Notes on Instrument Safety
carefully before installing and starting up
the device.
3.1 Installation Requirements
The following requirements apply to the installation of the device:
−Stable laboratory bench for microscope system on vibration-isolated base plate or vibration-isolating,
air-cushioned microscope stage
Power Requirements:
Two additional wall outlets are required to connect the system (VivaTome and HXP 120).
Ambient Conditions:
1. Operation, specified power T = 22 °C ±3 °C without interruption (24 h per day,
independent of whether the system is operating or
switched off)
2. Operation, reduced power T = 10 °C to 35 °C, any conditions different from item 1.
3. Warm-up time 1 h, for high-precision and/or long-term measurements
≥2 h
4. Relative humidity < 65 % at 30 °C
5. Operation altitude max. 2000 m
A constant temperature of 22 °C is ideal for an invariable and optim
um performance of the
system.

START-UP
Carl Zeiss Requirements as to the Microscope Stand VivaTome
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3.2 Requirements as to the Microscope Stand
VivaTome is permitted for operation in combination with the following microscope stands:
−Axio Observer (all variants)
−Axio Imager (all variants)
−Axio Examiner (all variants, except for 2TV zoom tube)
The manufacturer does not assume liability for any use that is unrelated to the
intended use.
Any use of VivaTome in combination with microscope stands having an order number other
than that indicated in the present manual is not in accordance with the intended use.
Set up the microscope vibration-free on a vibration-damped table or on an anti-vibration plate.
The optimum performance of the complete system can be achieved only with a 100/0
deflection to the camera port with VivaTome mounted.
Long-
term experiments require a stable ambient temperature to avoid any artifacts caused by
thermal expansion.
In addition to that, connection to the microscope requires a camera adapter 60N-C 1" 1.0x.

START-UP
VivaTome Basic Microscope Settings Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 17
3.3 Basic Microscope Settings
•Connect the microscope to the computer, if desired.
•If not yet done, install the image processing software AxioVision Rel. 4.8 or higher on the computer.
•As described in the respective operating manual, insert one of the indicated objectives into the
objective nosepiece and keep the Immersol 518 F®immersion oil ready to hand.
Read in any case the Immersol 518 F®safety data sheet.
3.4 Mounting VivaTome
The white light source must not be switched on until after having connected the output of the
liquid light guide tightly with VivaTome and VivaTome firmly screwed to the coupling point of
the microscope.
Before mounting VivaTome to the Axio
Observer side port it is necessary to screw two
supporting feet to it. See chapter 3.5.
•Remove the dust cap (Fig. 3-1/3) from VivaTome.
•Screw the c-mount camera adapter (Fig. 3-1/2) onto VivaTome (Fig. 3-1/1).
•If necessary (if using an Axio Observer), mount the supporting feet. See chapter 3.5
•Fasten the c-mount camera adapter (Fig. 3-1/2) to the microscope using a union ring.
Fig. 3-1 Mounting VivaTome to the microscope (here Axio Observer)

START-UP
Carl Zeiss Mounting Supporting Feet VivaTome
18 423639-9000-701 M60-2-0046 e 05/2012
3.5 Mounting Supporting Feet
Before mounting VivaTome to the Axio
Observer side port it is necessary to screw two
supporting feet to it.
•Unscrew the two setscrews (Fig. 3-2/1)
approximately half way from the supporting
feet (Fig. 3-2/2) using an Allen key SW 3.
•Hold the setscrews and screw them (Fig. 3-2/2)
into VivaTome as far as it will go.
•Now, screw the supporting feet in or out to the
required height.
•Turn the counternut (Fig. 3-2/3) downwards to
secure the supporting foot.
3.6 Mounting the Camera
The following cameras can be connected to the
scanhead:
−AxioCam MRm
−AxioCam HRm
All cameras are mounted in the same way.
•Loosen the clamping screw (Fig. 3-3/1) (Allen
key SW 3) and remove the camera adapter
(Fig. 3-3/2) obliquely frontward from the holder
(Fig. 3-3/3).
•Pull off the dust caps from the camera adapter
and the camera.
Fig. 3-2 Mounting the supporting feet
Fig. 3-3 Removing the camera adapter

START-UP
VivaTome Connecting VivaTome Carl Zeiss
M60-2-0046 e 05/2012 423639-9000-701 19
•Screw the camera adapter (Fig. 3-4/1) onto the
camera (Fig. 3-4/2).
•Introduce the camera adapter obliquely from
the front into the holder (Fig. 3-3/3). In doing it,
pay attention to the position of the camera
(horizontal position, cable above).
•Tighten the clamping screw (Fig. 3-3/1) using a
3 mm Allen key.
3.7 Connecting VivaTome
•Connect the VivaTome power supply unit with
a mains socket outlet.
•Connect the DC output of the power supply
unit to the socket (Fig. 3-5/2) on the back of the
device.
•Insert the liquid light guide into the opening
(Fig. 3-5/1) on the back of the device as far as it
will go.
•Tighten the securing screw (Fig. 3-5/4) using a
3 mm Allen key to secure the liquid light guide.
•Connect the USB cable to the USB connector
(Fig. 3-5/3).
Fig. 3-4 Screwing the camera adapter onto
the camera
1Connection for liquid light guide
2Connection for power supply unit
3USB connector
4Securing screw
Fig. 3-5 Connecting VivaTome

OPERATION
Carl Zeiss Control and Functional Elements VivaTome
20 423639-9000-701 M60-2-0046 e 05/2012
4OPERATION
For detailed information on the
operation of VivaTome, please see the manufacturer's
operating manual included in the delivery.
The present chapter describes only the operation of the device in connection with Zeiss
peripherals.
4.1 Control and Functional Elements
VivaTome has the following control elements:
−ON/OFF switch (Fig. 4-1/1)
−Grid pattern selection switch (Fig. 4-1/2)
Switch-over between the two grid patterns.
−Filter selection switch (Fig. 4-1/3)
Switch-over between different filters.
−Shutter pushbutton (Fig. 4-1/4)
To open/close the VivaTome shutter.
4.2 Switching On
•Switch on the microscope.
The reflector turret must be in its free passage position!
•Select a suitable filter module. See manufacturer's operating manual.
For changing the filter module see chapter 4.4.
•Switch on VivaTome using the ON/OFF switch (Fig. 4-1/1).
1ON/OFF switch
2Grid pattern selector
3Filter selector
4Shutter pushbutton
Fig. 4-1 Control and functional elements of
VivaTome