Cell Dynamics W8 User manual

ORIGINAL INSTRUCTIONS
ENGLISH / AMERICAN

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Table of Contents
1. Warnings and precautions................................................................................................................... 10
1.1. Intended use of the instrument .................................................................................................. 10
1.2. Limitations................................................................................................................................... 10
1.3. Required expertise of the operator............................................................................................. 11
1.4. Emergency stop procedure ......................................................................................................... 11
2. Safety labels, Hazards and Indicators.................................................................................................. 12
2.1. Safety instructions....................................................................................................................... 12
2.2. Delivery and Transportation........................................................................................................ 13
2.3. Area of Use and Positioning ........................................................................................................ 14
2.4. Electricity and fire hazards .......................................................................................................... 15
2.5. Electromagnetic compatibility (EMC).......................................................................................... 17
2.6. Acoustic emission ........................................................................................................................ 17
2.7. Liquid spillages and leakages....................................................................................................... 17
2.8. Temperature................................................................................................................................ 17
2.9. Mechanical hazards..................................................................................................................... 18
2.10. Chemical and Biological Hazards............................................................................................. 18
2.11. Service ..................................................................................................................................... 19
2.12. Disposal.................................................................................................................................... 20
3. Introduction and applications ............................................................................................................. 21
3.1. The W8 Physical Cytometer......................................................................................................... 21
3.2. Working principle ........................................................................................................................ 22
3.3. Applications ................................................................................................................................. 22
4. Schematic overview of the W8 Physical Cytometer............................................................................ 23
4.1. Component list ............................................................................................................................ 27
5. Installation of the W8 Physical Cytometer.......................................................................................... 28
5.1. Setting Up the W8 Physical Cytometer ....................................................................................... 28
5.2. PC requirements.......................................................................................................................... 28
5.3. LIBRA: software installation......................................................................................................... 28
6. Operative protocols............................................................................................................................. 29
6.1. Starting Up the W8 Physical Cytometer...................................................................................... 29
6.2. Filter connection.......................................................................................................................... 29

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6.3. Waste bottle positioning and connection ................................................................................... 30
6.4. W8 caps positioning .................................................................................................................... 30
6.5. Reservoirs: preparation and connection..................................................................................... 31
6.6. W8 chip information and insertion procedure............................................................................ 34
6.7. Cleaning Solution preparation..................................................................................................... 35
7. LIBRA: User Manual............................................................................................................................. 37
7.1. Experiment Settings..................................................................................................................... 37
7.2. Sorting Options............................................................................................................................ 40
7.3. Operative panel........................................................................................................................... 41
7.4. Control panel: FILL....................................................................................................................... 42
7.5. Control panel: MEASURE............................................................................................................. 44
7.6. Control panel: CLEAN................................................................................................................... 47
8. Troubleshooting: a short guideline on possible issues........................................................................ 51
8.1. Incompatible number and size of samples.................................................................................. 51
8.2. Out-of-scale liquids volumes ....................................................................................................... 51
8.3. Unsatisfactory W8 Chip status and positioning........................................................................... 51
8.4. Incorrect tubes positioning.......................................................................................................... 52
8.5. Incorrect naming of the LIBRA folder.......................................................................................... 52
8.6. Air Bubbles in the W8 Chip channel ............................................................................................ 52
8.7. Multiple samples in the W8 Chip channel................................................................................... 53
8.8. Impurities in the W8 Chip channel .............................................................................................. 53
8.9. Switching to a new sample.......................................................................................................... 53

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Copyright © 2021 CellDynamics. All rights reserved.
No part of this User Manual may be reproduced, stored in a retrieval system, transmitted, published, or distributed in
any form or by any means, electronically, mechanically, by photocopying, microfilming, recording, or otherwise, without
the prior written consent of CellDynamics.
However, notwithstanding the foregoing, the owners of the W8 Instrument may make copies solely for purposes of
training personnel in the use of the unit within their business or organization.
Maximal care has been taken by CellDynamics in the preparation of this User Manual. However, CellDynamics shall not
be liable for any technical or editorial errors or omissions contained herein or for incidental or consequential damages
in connection with the furnishing, performance, or use of this document. The information in this document are provided
“as is” without warranty of any kind, and they are subject to change without notice.
All other trademarks mentioned in this document are the property of their respective owners and are used for
identification purposes only. Unless otherwise specifically indicated, CellDynamics products and services are for
research use only and not for therapeutic or diagnostic use.
Revision Date:
June 8th, 2021

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Limited warranty
● The W8 Physical Cytometer Unit is warranted for 12 months from the date of delivery.
● The appropriate use of the device is specified in the W8 Physical Cytometer User Manual.
● If damage is due to the misuse of the W8 Physical Cytometer, all warranties shall be void. This includes
damage to the device itself as well as any consequential damages.
● Warranties are void if the damage is due to the use of analysis solutions that are not produced and/or
distributed by CellDynamics or its official commercial partners and/or distributors. This includes damage to
the device itself as well as any consequential damages.
● Under all circumstances, liability is limited to the value of the W8 Physical Cytometer. Any liability beyond
the value is excluded.
If the product proves defective during the warranty period, CellDynamics could decide one of the following
options:
(1) provide specialized technicians to repair the product at no charge for parts or labor;
(2) replace the product with a comparable product which may be new or refurbished;
(3) refound the re-evaluated value of the product based on the paid amount and the product condition.
To obtain service under this warranty the Customer must notify CellDynamics Support or its authorized
service representative of the encountered defect before the expiration of the warranty period.
In the maintenance of the product, CellDynamics may use new or equivalent-to-new parts, assemblies or
products for equal or improved quality. All defective parts, assemblies, and products become the property
of CellDynamics.
CellDynamics may require the return of parts, assemblies and products to a designated CellDynamics Depot
or the CellDynamics representative from which the part, assembly, or product was originally purchased.
Return and claims will be handled according to the current CellDynamics procedure.
These warranties shall not apply to any defect, failure or damage caused by improper use or inadequate
maintenance and care, done by personnel not authorized by CellDynamics.

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EC DECLARATION OF CONFORMITY
Referring to the Directive 2006/42/CE II.A on machinery, and amending Directive
DECLARATION IN THE ORIGINAL LANGUAGE
We: CellDynamics i.s.r.l.
Via Villa Clelia 62/d
40026 IMOLA (BO) - ITALIA
as manufacturer of the product, declare that the machine:
Physical cytometer for the analysis of mass density, size, shape of samples comprised between
50 and 500 μm:
MODEL : __W8 Physical Cytometer______
REFERENCE CODE: __W8-
FABRICATION YEAR: ___2020______
to which this declaration refers complies with the following legislative provisions:
- DIRECTIVE 2006/42/EC OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 17
May 2006 on machinery, and amending Directive 95/16/EC (recast)
- DIRECTIVE 2014/30/UE OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 26
February 2014 on the harmonization of the laws of the Member States relating to electromagnetic
compatibility (recast)
- DIRECTIVE 2011/65/UE OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 8
June 2011 on the restriction of the use of certain hazardous substances in electrical and electronic
equipment (recast)
The machine also complies with the provisions of the following harmonized standards :
EN ISO 12100:2010
Machinery safety - General design principles - Risk assessment and risk reduction
EN 61326-1:2013
Electrical equipment for measurement, control and laboratory - Electromagnetic
compatibility requirements

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1. Warnings and precautions
The instrument is conceived for safe use when the installation and operation procedures are
executed properly and in accordance with the instructions described in the present User Manual.
This section describes potential risks related to the installation and usage of this instrument and its software.
Moreover it provides guidelines and crucial safety information with the aim to avoid and/or minimize such
risks.
Follow the instructions carefully to prevent potential dangers and to protect yourself, others, and the
equipment. This instrument must be used only as specified by the manufacturer.
The safety instructions described in this document are considered as an implementation of the laboratory
policies, and local working area safety directives, in which the instrument is located, as well as standards for
health, safety and prevention of accidents.
Local authorities can provide safety details on the installation and operation of electrical power supply,
building constructions and maintenance for the correct placement of the device.
The W8 Physical Cytometer is for Research Use Only, not for use in Diagnostic Procedures.
A summary of the main concepts and procedures described in this User Manual, including the most
common issues that the operator may encounter while using the instrument (Troubleshooting), is available
on the attached QUICK START GUIDE document.
1.1.Intended use of the instrument
The W8 Physical Cytometer is conceived and tested for measuring the mass density, size and weight of
sphere-like samples (i.e. spheroids, organoids, organic or hybrid microparticles, etc.). Moreover, the
instrument is able to discriminate such samples based on the selected mass density or diameter range by the
operator. The instrument can operate for samples having sizes comprised between 50 and 500 micrometers
(μm) in diameter. Samples must be handled, introduced into the instrument, measured and sorted by
following the procedures described in this User Manual.
1.2.Limitations
DO NOT use the instrument for samples outside of the indicated size range of 50 - 500 μm in diameter.
Samples bigger than 500 μm may cause clogging of the fluidic system and lead to instrument malfunctions.
Samples smaller than 50 μm will prolong the analysis time and they are not accurately detectable. Moreover,
they may persist into the fluidic system even after the cleaning protocol, leading to biological contaminations
overtime.
DO NOT use nanosized materials or materials that can interact with Silicon or Poly(methyl methacrylate)
(PMMA). This can ruin or contaminate the internal fluidic system, as well as creating malfunctionsor leakages
over time.

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DO NOT use other liquids except for the CellDynamics W8 Analysis Solution, the Cleaning Solution and
Milli-Q water. The CellDynamics W8 Analysis Solution is specifically conceived to allow the instrument to
operate properly and maintain its accuracy. The Cleaning Solution must be prepared as indicated in Section
6.5.
Use the correct number of samples inside the Sample Holder. This is key for the successful outcome of the
experiment. Follow the suggested values in Section 6.5.
DO NOT use the instrument when the temperature of the environment is outside the 15 - 35 °C range. For
maximum accuracy, it is suggested to operate within 18 and 30 °C.
DO NOT incline the centrifuge tube, or attempt the shaking of the solution by hand. The W8 Caps are not
sealed, and this would lead to a spillage of the solution containing the samples. Moreover, to avoid possible
contaminations, never allow the solution to be in contact with the W8 Caps.
1.3.Required expertise of the operator
The W8 Physical Cytometer is accessible to all researchers, regardless of their technical skills.
Not specialized personnel can operate the instrument by simply following the instructions. Due to its
simplicity of use, no specific skills are quired as long as the operator has knowledge of the purpose of his
research, as well as of the biological material employed and its hazards.
This makes the instrument suitable and safe for research purposes even when supervising young students.
Moreover, after purchasing the instrument, a demo and training session with a CellDynamics specialist will
be scheduled to make the operator in the best position to work consciously and safely.
1.4.Emergency stop procedure
The instrument is designed to be stable under the conditions described in this User Manual.
However, in case of an error made by the operator (i.e. centrifuge tube not connected), or an anomaly of the
system, quickly click the “Pause” button on the software.
Most importantly, in the case of an emergency, the operator must stop the procedure and turn the
instrument off. Emergency reasons can range from the spillage of harmful materials to external factors such
as laboratory fire alarms.
If an emergency occurs while measuring, follow the Emergency Stop procedure below:
1) Click the “Pause” button on the software.
2) Switch off the instrument from the power button on the rear panel.
3) Unplug the main power source cable.
NOTE: If an emergency occurs during the experiment setup procedure, simply switch off the instrument from
the power button on the rear panel and unplug the main power source cable.

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2. Safety labels, Hazards and Indicators
The following signals are present throughout this User Manual and are associated with information related
to specific risks or hazards as reported below:
Indicates a potentially hazardous situation, which, if not avoided, could result in serious
injuries or death.
Indicates a potentially hazardous situation, which, if not avoided, may result in minor or
moderate injuries. It may also be used to alert against unsafe practices.
General risk of danger. Every sentence, picture or part associated with this symbol in this
document needs to be consulted carefully.
Risk of electric shock.
Risk of fire.
Risk due to liquid spills or corrosion.
Risk from biological hazards or harmful chemicals.
Risk of injuries due to wrong handling and transportation, or risk for the instrument during
transportation.
2.1.Safety instructions
The safety of the instrument, and its operations, are no longer guaranteed if the instrument is not
working properly. If this happens, execute the Emergency Stop procedure, then contact the CellDynamics
local service provider for technical support.
The instrument is intended for indoor use only.
In the case of analyzing or operating with hazardous or unknown materials/substances the
instrument must be placed and used under a Class II / III Biological Safety Cabinet.
Stop using the instrument and unplug it from the main power source if:
it has been opened or disassembled
it has been dropped or damaged
it has damaged or broken parts
it has a damaged power cable

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an object has entered through the ventilation slots
an object has been dropped into the instrument
an internal spillage occurred
2.2.Delivery and Transportation
The instrument, and its accessories (listed in Section 4.1), are shipped into a protective wooden case
specifically designed. Based on the customer's request, the case could be also shipped securely positioned
onto a wooden pallet. The packaging is provided by CellDynamics and it must be used uniquely.
Inspection of the delivery. Carefully inspect the entire packaging upon receipt of the W8 Physical Cytometer.
In case of signs of mishandling or damage, file a claim with the carrier immediately. If the shipment is
separately insured, file a claim with the insurer.
For proper and secure handling, the case includes two handles, positioned on two opposite sides, as
graphically described in Figure 2.1.
In case the pallet is included in the shipment, the transportation of the entire packaging (case and pallet),
from the collection center to the laboratory, must occur using a trans-pallet. Once reached the location, the
wooden case can be lifted and repositioned by hand. Differently, when the instrument is shipped without
the pallet (wooden case only), it can be transported by hand.
FIGURE 2.1 GRAPHICAL REPRESENTATION OF THE WOODEN CASE AND PALLET
Always keep the instrument vertically, also during transportation.

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The vertical position is achieved by orienting the instrument in a manner that the upper part of it,
represented by the “UP” arrow in Figure 2.2, remains upwards and orthogonally with respect to the
horizontal plane. The instrument must be maintained vertical at any moment. Never incline the instrument
on any side for any reason. A bubble level, on the top panel, allows a reference point to maintain the correct
inclination (see Figure 4.2 Section 4).
FIGURE 2.2 W8 PHYSICAL CYTOMETER ORIENTATION
Transport the instrument with care.
Dropping the instrument can may cause damages to it. This may occur even when the instrument is packed.
Moreover, excessive vibrations can cause malfunctions.
In the case of shipping the instrument back to CellDynamics, carefully perform the CLEAN (Section 7.6)
procedure for decontamination before shipment. If in doubt, request the CellDynamics assistance for the
decontamination protocol and/or shipment procedure.
Always handle and transport the instrument following the safety guidelines for heavyweight
transportation in order to prevent injuries.
2.3.Area of Use and Positioning
The W8 Physical Cytometer is designed to be positioned onto a fixed laboratory benchtop.
Its highest performance can be reached when placed onto an anti-vibration support (i.e. Atomic Force
Microscopy benchtop). The device should not be located onto a normal table or any surface or support in
close proximity of a source of vibration (i.e. pumps, sonic baths etc.).

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In the case of analyzing or operating with hazardous or unknown materials/substances, the instrument
must be placed and used under a Class II / III Biological Safety Cabinet.
Do not use the instrument in a wet location or environment. Particularly, do not place the instrument on
areas with high humidity.
Maintain at least 15 cm of distance between the W8 Physical Cytometer and any wall, object or PC. This
distance is also needed for proper ventilation as discussed in Section 2.8.
Moreover, keep a free area in front of the instrument to allow the opening of the safety door for proper use
(Figure 2.3).
FIGURE 2.3 SCHEMATIC REPRESENTATION OF THE POSITIONING OF THE INSTRUMENT FOR SAFETY AND PROPER VENTILATION:
DISTANCES FROM WALLS,OBJECTS AND/OR PC.
The main switch button of the instrument and the plug of the power cable must be easily accessible to the
operator. This is crucial in case of emergency when the instrument needs to be turned off and the power
source needs to be unplugged.
2.4.Electricity and fire hazards
The device satisfies all applicable requirements of the DIRECTIVE 2006/42/EC OF THE EUROPEAN
PARLIAMENT AND OF THE COUNCIL of 17 May 2006 on machinery, which includes all the safety requirements
concerning the presence of dangerous electrical voltage.

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However, it is important to highlight that:
Electrical devices may arise potential risks like electric shock, electric short and overheating. Events
like electric shock may lead to severe personal injury or death.
An electric short and overheating may ignite fire. This is due to the possible production of sparks
which, in contact with combustible vapors or flammable materials can generate explosion or fire.
To minimize the hazard of an electric shock, handle fluids with superior attention when in proximity
of the instrument. Caution must be taken to avoid fluids entering into the device case. In case of spills, always
unplug the main power first and clean immediately.
Do not place and/or use the instrument in hazardous classified areas such as combustible vapors
environments. In the event of visual fire or smoke quickly perform the Emergency Stop procedure if
possible. Subsequently, contact the CellDynamics technical support.
Do not remove and/or modify any cover protection or part of the case.
Never insert an external object through a fissure or opened part of the instrument.
The instrument is conceived to minimize the risk of electric shock, electric short, and the spread of fire. Only
authorized CellDynamics technicians are allowed to remove, modify or manipulate covers or internal parts
of the instrument.
Connect the device to the power supply with the provided power cable only. No third-party power
suppliers cables are allowed. Follow the mounting instructions for the instrument in Section 5.1 to ensure
the proper contention.
Periodically verify that the cable meets the electrical specifications required by the safety regulation. If the
electrical safety check is not satisfied contact CellDynamics.
The instrument is equipped with a grounding-type plug for safety. Do not try to insert the plug into a non-
grounded power outlet. If you cannot insert the plug into the outlet, contact your local electrician to replace
the outlet.
Do not use extension cables or power strips. Do not overload the electrical outlet.
The main power supply source must be visible and accessible at any time to be rapidly unplugged in case
of danger.
A computer, or laptop, is the only peripheral device that can be connected to the W8 Physical Cytometer.

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2.5.Electromagnetic compatibility (EMC)
The instrument satisfies the electromagnetic compatibility (EMC) regulation, based on the
DIRECTIVE 2014/30/UE OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 26 February 2014 on the
harmonization of the laws of the Member States relating to electromagnetic compatibility (recast).
2.6.Acoustic emission
The A-weighted sound pressure level (not-continuous), that the instrument generates in the workplace, is
less than 70 dB(A).
2.7.Liquid spillages and leakages
Always examine the visible tubing sets, reservoirs, bottles and their closures, as well as the injection system
before switching the instrument on and before launching the measurements. Particular attention should be
focused on the correct position of the small o-rings inside the Pin at the top of the W8 Caps (Section 6.4).
Take special care while handling liquids.
Avoid liquids entering into the instrument. This would avoid the risk for electric shorts, electric shock, or fire.
The risk of accidental spillages and splashes must be minimized by protecting the instrument. In the
unfortunate case of spillage, unplug the instrument from the power source and clean up immediately.
For cleaning, do not use cleaning agents in liquid or aerosol form, use a slightly damped cloth only.
Stop operating the instrument if liquids have accidently entered into the instrument.
If detecting a leakage related to the fluidic network, perform the Emergency Stop procedure. If the leakage
is accessible, clean and dry it. If the spillage involves the internal fluidic system, do not remove the covers
and contact the CellDynamics Team to perform the technical service.
2.8.Temperature
Keep the instrument at room temperature.
The instrument is tested to perform between 15 and 35 °C. However, for maximum accuracy, it is suggested
to operate within 18 and 30 °C.

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Protect the instrument from cold conditions.
In the case of a drop in temperature of the environment where the device is located (i.e. 5 °C), consider that
this may have caused internal condensation. In such cases, transport the device to room temperature
condition and wait for the instrument to dehumidify before using it.
Protect the instrument from overheating.
Do not use the instrument when the temperature condition exceeds 35°C. Keep the instrument away from
direct sunlight and ensure adequate air circulation in the room where the instrument is placed and in
proximity of the instrument.
Air circulation must be guaranteed by always maintaining a space of at least 15 cm on all sides of the
instrument when operating. Avoid any source of heat in proximity of the instrument (i.e. radiators, stoves,
or any equipment that generates heat).
2.9.Mechanical hazards
Moving and/or rotating parts are potential mechanical hazards. Do not open the main case
or the front and rear panels. The instrument includes pumps and tubing which, if disconnected and wrongly
activated during the operations, may lead to spillages. The main case, panels and front door of the instrument
are conceived to protect the operator. Defective or inadequate safety equipment has to be considered
hazardous and it needs to be replaced.
Do not disconnect or adjust in any manner the fluidic network, nor evade safety measures, during operation.
Always stop a procedure (Pause button), before handling hardware accessories such as centrifuge tubes, W8
analysis solution or waste, except when it is part of the procedure indicated by the software.
Take special care when connecting the sample tubes by carefully following the procedure in Section 6.3.
LIGHT SOURCE: The instrument is equipped with a 3 V white LED light. Although this light source is not
considered dangerous it is suggested to not stare directly at the light source for a long period of time.
2.10. Chemical and Biological Hazards
Chemical and biological hazards
Chemical and biological substances employed while operating the instrument may cause contamination
and/or infection which may lead to severe personal injury or death.
All chemical, biological and/or clinical samples must be considered potentially dangerous or infectious.
The operator must select and wear personal safety equipment according to the SDS and/or MSDS document
of the substance, solution or product used.
This must be accomplished to prevent any form of risk, ranging from hazardous, toxic, acidic or radioactive
chemicals, to carcinogenic substances that may be present into the sample of analysis.

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Even when not highlighted on the SDS of the substances used, wear protective gloves and clothing, as well
as safety glasses to prevent contact of the substance with skin and eyes. This is highly recommended for
state-of-the-art research when no side effects are known.
In the case of analyzing or operating with hazardous or unknown materials/substances, the instrument
must be placed and used under a Class II / III Biological Safety cabinet.
DO NOT continue to handle contaminated accessories or parts of the instrument. The instrument must be
carefully decontaminated if hazardous material has been used or spilled (follow the “Sterilize” protocol
described in Section 7.6).
If operating with volatile substances or aerosols that may leak from the tubes, protect the mouth and nose
adequately.
The waste must be considered hazardous and it must be handled following universal laboratory
precautions. Local laboratory regulations must be adopted for the disposal procedure of the waste.
Tubes and consumable materials in contact with hazardous substances must be decontaminated before
disposal.
Parts that contain chemical or biological hazardous materials or liquids, or parts that have been in contact
with such materials, should be considered as hazardous and treated accordingly. This also applies to failure
parts that need to be substituted.
2.11. Service
Do not service the instrument yourself.
Inappropriate servicing of the instrument may create hazards to users, lead to instrument malfunctions or
damages, causing unpredictable results, and reducing the instrument’s life.
Unauthorized service will void the warranty.
Always call the CellDynamics Team for proper service. Only CellDynamics technicians, or trained technicians
delegated by CellDynamics, can proceed with the inspection, service and repair of the instrument.
Do not switch the instrument on if parts are not working properly or they need to be replaced.
If replacements are not available, unplug the instrument from the main power source and contact the
CellDynamics Team.
In case the replacement of one or more parts is needed, ensure that only approved components are used.
These can be provided by CellDynamics or by specifically selected third parts. Unapproved components can
cause malfunction and/or damage of the instrument as well as unpredictable and untrustworthy results.
CellDynamics is not responsible for instrument malfunctions and/or damages caused by not approved
components used for replacements of original parts.
The use of unapproved components will void the warranty.
The authorized CellDynamics service provider should perform all the safety checks at the end of the repair
procedure to ensure that the device operates properly.

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Upgrades
Only use options and upgrades recommended by CellDynamics or delegate representatives. These can be
related to both the instrument and the software.
Information can be gathered on the website www.celldynamics.it, or contacting the CellDynamics Tech Team
at [e-mail].
2.12. Disposal
When at the end of the instrument’s life, adopt the WEEE (Disposal Waste of Electrical and Electronic
Equipment) regulation and legislation which may be different based on the country.
If the instrument, and/or parts of the equipment, are contaminated, dispose them according to the WEEE
compliance schemes, and in line to the designated facility center for your area.
This will help to preserve natural resources and human health.
Contact CellDynamics for the evaluation of the end-of-life service.