ChemoMetec NucleoCounter NC-202 User manual

NucleoCounter®NC-202™
Instrument User Guide
P/N 991-2020
Revision 1.1
November 2019
ChemoMetec A/S
Gydevang 43 · DK-3450 Allerød · Denmark
Telephone: [phone]
Website: www.chemometec.com
E-mail: [e-mail]

991-2020 Rev. 1.0 May 2019
ii
Caution!
This equipment must be operated as described in this user guide and documents referenced herein. Please
read the entire guide and referenced documents before using the NucleoCounter® NC-202™. Gloves or
protective clothing may not be shown in the pictures in this user guide; however, ChemoMetec recommends
that the user wear suitable protective clothing.
Contacting Support
Technical information including product literature, answers to questions not covered in this document and
referred documents regarding the operation of the NucleoCounterNC-202™is available from our support
team:
•E-mail questions to [e-mail]
•To speak with a Technical Support Specialist, call ([phone]
Please create support files in NC-View™via Help →Create support files, before contacting us.
Sales and Ordering Information
To order the NucleoCounterNC-202™, NC-View™software and consumables, call ([phone], fax (+45)
48 13 10 21, or e-mail [e-mail].
Disclaimer Notices
The material in this document and referred documents is for information only and is subject to change without
notice. While reasonable efforts have been made in preparation of these documents to ensure their accuracy,
ChemoMetec assumes no liability resulting from errors or omissions in these documents, or from the use of
the information contained herein.
ChemoMetec reserves the right to make changes in the product design without reservation and without
notification to its users.
Copyright Notices
Copyright © ChemoMetec A/S 2019. All rights reserved including graphics and images. No part of this
publication, software and referred documents may be reproduced, stored in a retrieval system, or transmitted
in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, without the prior
written consent of ChemoMetec A/S, Gydevang 43, DK-3450 Allerod, Denmark.
ChemoMetec® and NucleoCounter® are registered trademarks owned by ChemoMetec. NucleoCassette, Via2-
Cassette and NC-View are trademarks of ChemoMetec.
All other trademarks are the property of their respective owners.

Declaration of Conformity
991-2020 Rev. 1.1 November 2019 iii
Declaration of Conformity
Name of product: NucleoCounterNC-202™
Type: Automated Cell Counter
Other identifying data: Part no. 900-2020
We declare under our sole responsibility that the products, to which this declaration
relates, are in conformity with the Council Directives on the approximation of the laws of
the EEC Member States relating to the following:
Directives
Electromagnetic Compatibility (2014/30/EU) EMC
Low Voltage Directive (2014/35/EC) LVD
Machinery Directive (2006/42/EC) MD
Standards
EMC:
EN 61326-1:2013 Electrical equipment for laboratory use
EN 61000-3-2: 2014 Limits for harmonic current emissions
EN 61000-3-3: 1995 +A1 +A2: 2013 Limitation of voltage changes,
voltage fluctuations and flicker
Standards
MD:
EN 12100-1: 2003 Safety of machinery; Basic concepts, general
principles for design —Part 1
EN 12100-2: 2003 Safety of machinery; Basic concepts, general
principles for design —Part 2:
EN ISO 14121-1:2007 Safety of machinery; risk assessment
This device complies with Part 15 of the FCC Rules. Operation is subject t o the following
two conditions: i) this device may not cause harmful interference; and ii) this device must
accept any interference received, including interference that may cause undesired
operation.
Date: 2019-April-12th Signed:
Name: Eva Louise Lange
Position: Quality Manager
Name and address of manufacturer:
ChemoMetec A/S
Gydevang 43
DK-3450 Allerod
DENMARK
www.chemometec.com

WEEE Directive Information –Europe Only
991-2020 Rev. 1.1 November 2019 iv
WEEE Directive Information –Europe Only
Correct Disposal of This Product
(Waste Electrical & Electronic Equipment) - Europe only
This marking shown on the product or its literature, indicates that it should
not be disposed of together with other household wastes at the end of its
working life. To prevent possible harm to the environment or human health
from uncontrolled waste disposal, please separate this from other types of
wastes and recycle it responsibly to promote the sustainable reuse of
material resources.
Business users should contact their supplier and check the terms and
conditions of the purchase contract. This product should not be mixed with
other commercial wastes for disposal.
This information is listed in “Appendix A: WEEE Directive Information in
More EU Languages”.

Introduction and Intended Use
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Introduction and Intended Use
The NucleoCounter® NC-202™is a compact, automated cell counter intended for cell
counting within the life science and pharmaceutical markets. The NucleoCounter®
NC-202™system is intended for research use only, not for diagnostic use. The
NucleoCounter® NC-202™uses disposable cassettes from ChemoMetec.
The NucleoCounter® NC-202™is an advanced cell counter based on low magnification
fluorescence microscopy using LED light and CMOS camera technology, combined with
automated image analysis. Cells are analyzed using two fluorescence channels, where the
spatial position is maintained between images, allowing direct comparison and
quantification of parameters at single cell level. Low magnification (×2.1) and a large
sensor area (2/3”) facilitate a high volume of analytical samples, thereby providing great
statistical performance. The NucleoCounter® NC-202™ light sources deliver homogeneous
sample illumination, which reduces systematic errors in fluorescence signal intensities
across the measurement area and increases the accuracy of cell counting values.
The NucleoCounter® NC-202™ requires NC-View™software to operate. NC-View™is
distributed with a list of protocols employing two fluorescence channels to analyze cells.
Instrument qualification protocols are included in the software.
The NucleoCounter® NC-202™ uses Via2-Cassettes™, which contain fluorescent dyes and
allow the NucleoCounter® NC-202™ to detect cells. The NucleoCounter® NC-202™is the
third generation of NucleoCounter® cassette instruments, succeeding the NucleoCounter®
NC-100 and NucleoCounter® NC-200™.
The NucleoCounter® NC-202™ChemoMetec part number is 900-2020.

Warnings and Precautions
991-2020 Rev. 1.1 November 2019 vi
Warnings and Precautions
Whenever the symbol appears on the NucleoCounter® NC-202™instrument, it
indicates that the manual must be consulted for precautions and warnings.
Power Supply and Cables
Use the shielded USB cable supplied with the NucleoCounter® NC-202™to ensure that
appropriate EMI classification is maintained for the intended environment.
The USB interface connector must only be connected to SELV circuits. Ext ernal
computing devices connected to the USB interface connector of the NucleoCounter® NC-
202™must comply with standards UL 1950 and IEC/EN 60950.
Table 1: Description of the NucleoCounter® NC-202™ USB 3.0 micro interface connector
The NucleoCounter® NC-202™is powered by an external 24VDC power supply. For safe
use, please follow the instructions for connecting the power supply.
The NucleoCounter® NC-202™should only be used with an external power supply
purchased from ChemoMetec.
The detachable DC power supply cable set and the appliance inlet of the external
power supply are considered as the disconnecting device.
The main supply cable and plug of the external power supply should comply with any
national regulations.
The user is made aware that if the NucleoCounter® NC-202™and the external power
supply are used in a manner not specified in this user guide, the protection may be
impaired.
1
In normal operation mode (refers to Pin no 5 and/or 8)
Pin
Pin Name
Pin Description
Maximum
Voltage level1
1
VBUS
+5V
+5 VDC
2
USB D-
USB 2.0 differential pair
+3.5 VDC
3
USB D+
USB 2.0 differential pair
+3.5 VDC
4
OTG ID
USB OTG ID for identifying lines
5
GND
Ground
0 VDC
6
USB3 SSTX-
USB 3.0 signal transmission line
7
USB3 SSTX+
USB 3.0 signal transmission line
8
GND
Ground
9
USB3 SSRX-
USB 3.0 signal receiving line
10
USB3 SSRX+
USB 3.0 signal receiving line

Warnings and Precautions
991-2020 Rev. 1.1 November 2019 vii
Electromagnetic Interference
NOTE: This equipment has been tested to meet the Class B digital device limits, pursuant
to Part 15 of the FCC Rules. These limits are designed to provide reasonable protection
against harmful interference in a residential installation. This equipment generates, uses
and can radiate radio frequency energy and, if not installed and used in accordance with
the instructions, may cause harmful interference to radio communications. However, there
is no guarantee that interference will not occur in any given installation. If this equipment
does cause harmful interference to radio or television reception, which can be determined
by turning the equipment off and on. The user is encouraged to try to correct the
interference by one or more of the following measures:
•Reorient or relocate the receiving antenna
•Increase the separation between the equipment and receiver
•Connect the equipment to an outlet on a circuit different from the receiver
connection
•Consult the dealer or an experienced radio/TV technician for help
Caution! Changes or modifications not expressly approved by the party responsible for
compliance could void the user's authority to operate the equipment.
Cassettes, Reagents, Solutions and Dispensers
With respect to the use and handling of cassettes, reagents, solutions and dispensers,
please refer to appropriate package inserts for these items.
Caution! Bottle-top dispenser: To protect against accidental spills, protective clothing, eye
protection and gloves must be worn when using potentially hazardous liquids.
General Precautions
Any biological specimen should be handled as if capable of transmitting infectious disease s
and disposed of with proper precautions according with national, federal, state and local
regulations.
Avoid specimen contact with skin or mucous membranes.
Never pipette by mouth.
Avoid cross contamination of samples during preparation. This can compromise the quality
of results.

Table of Contents
991-2020 Rev. 1.1 November 2019 1
Table of Contents
DECLARATION OF CONFORMITY .......................................................................................III
WEEE DIRECTIVE INFORMATION –EUROPE ONLY ..............................................................IV
INTRODUCTION AND INTENDED USE .................................................................................V
WARNINGS AND PRECAUTIONS ........................................................................................VI
1INSPECTION, UNPACKING AND INSTALLATION OF EQUIPMENT .................................... 3
1.1 INSTALLATION AND OPERATION QUALIFICATION OF NUCLEOCOUNTER®NC-202™ .................. 3
2CELL COUNTING WITH NUCLEOCOUNTER® NC-202™.................................................... 4
2.1 THE NUCLEOCOUNTER®NC-202™ INSTRUMENT ............................................................ 4
2.1.1 Fluorescence microscope .............................................................................. 4
2.1.2 Protocols ..................................................................................................... 4
2.1.3 The RUN button and LED indicator ................................................................ 5
2.1.4 Cassette fixture ............................................................................................ 5
2.1.5 USB 3 connection ......................................................................................... 5
2.2 THE VIA2-CASSETTE™.............................................................................................. 6
2.2.1 Loading the Via2-Cassette™ cassette ............................................................ 6
2.2.2 Via2-Cassette™ volume calibration dot code ................................................. 7
2.2.3 Cassette handling ......................................................................................... 7
3NUCLEOCOUNTER® NC-202™ – INSTRUMENT CONTROLS ............................................. 8
3.1 INTERACTIVE CONTROLS ............................................................................................ 8
3.2 NUCLEOCOUNTER®NC-202™ CONNECTIONS ................................................................ 9
3.2.1 NC-202™ power connection .......................................................................... 9
3.2.2 USB connection and initialization ................................................................ 10
4OPERATION OF NC-VIEW™ SOFTWARE AND NUCLEOCOUNTER® NC-202™ INSTRUMENT
11
4.1 NC-VIEW™SOFTWARE ........................................................................................... 11
4.2 PROTOCOL EXECUTION ........................................................................................... 11
4.2.1 Piston movements ...................................................................................... 12
4.3 PROTECTION OF THE OPTICAL SYSTEM ........................................................................ 12
5MAINTENANCE OF NUCLEOCOUNTER® NC-202™ ....................................................... 13
5.1 CLEANING ........................................................................................................... 13
5.1.1 Instrument cabinet ..................................................................................... 13
5.1.2 Cassette fixture .......................................................................................... 13
5.1.3 Contamination of the optics........................................................................ 13
5.1.4 Decontamination ........................................................................................ 13
6TROUBLESHOOTING FOR NUCLEOCOUNTER® NC-202™ .............................................. 14
6.1 FIRMWARE AND SOFTWARE VERSION.......................................................................... 14
6.2 POWER-ON FAILURE............................................................................................... 14

Table of Contents
991-2020 Rev. 1.1 November 2019 2
7TECHNICAL SPECIFICATIONS ..................................................................................... 15
7.1 NUCLEOCOUNTER®NC-202™ ................................................................................. 15
7.2 SAMPLE MEDIA .................................................................................................... 15
7.3 EU STANDARDS .................................................................................................... 15
8EQUIPMENT AND ACCESSORIES ................................................................................ 17
9OTHER DOCUMENTATION ........................................................................................ 17
APPENDIX A: WEEE DIRECTIVE INFORMATION IN MORE EU LANGUAGES ........................... 18
10 GENERAL DISCLAIMER .......................................................................................... 21

1 Inspection, Unpacking and Installation of Equipment
991-2020 Rev. 1.1 November 2019 3
1Inspection, Unpacking and Installation of
Equipment
Upon receiving the order from ChemoMetec, the box or boxes should be carefully
inspected for any damage that may have occurred during shipping. Any damage must be
reported to the carrier and to ChemoMetec immediately.
Unpack the boxes, saving the packing materials for possible later use.
Verify that the ChemoMetec packing list received refers to the correct and ordered
materials, and that nothing is missing.
If any part of the order was damaged during shipping, is missing or fails to operate, please
contact ChemoMetec immediately.
1.1 Installation and Operation Qualification of NucleoCounter® NC-202™
The NucleoCounter® NC-202™Installation Qualification (IQ) / Operational Qualification
(OQ) / Zero Count Application Note (# 994-2022) describes the IQ/OQ/Zero Count protocol.
Execution of the IQ protocol will ensure that the instrument is installed correctly. The
Operational Qualification (OQ) protocol verifies that the instrument is operating correctly.
Once IQ, OQ and Zero Count protocols are finalized successfully, the NucleoCounter® NC-
202™is ready for use.
The application note for the IQ/OQ/Zero Count protocol is available at
www.chemometec.com.

2 Cell Counting with NucleoCounter® NC-202™
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2Cell Counting with NucleoCounter® NC-202™
The NucleoCounter® NC-202™is a dual-color fluorescent cell counting instrument. The
NucleoCounter® NC-202™ is simple to operate, requiring little user input to execute e.g.
cell counting protocols.
The NucleoCounter® NC-202™ is operated by NC-View™ software.
2.1 The NucleoCounter® NC-202™instrument
The NucleoCounter® NC-202™instrument is shown below:
2.1.1 Fluorescence microscope
The NucleoCounter® NC-202™is a fully automated fluorescent microscope with a ×2.1
magnification. It is equipped with two light sources and a dual-band emission filter.
Together with the Via2-Cassette™the NucleoCounter® NC-202™provides a straight-
forward set-up, performing advanced analysis in an easy-to-use manner, so that personnel
training will be simple and variation between operators will be limited.
2.1.2 Protocols
The NucleoCounter® NC-202™ and NC-View™software are delivered with a list of protocols
to count cells and perform instrument validation. Currently, the protocols included are:
•Count & Viability
•NC-202 IQ/OQ

2 Cell Counting with NucleoCounter® NC-202™
991-2020 Rev. 1.1 November 2019 5
New protocols are developed at an on-going basis and an up-to-date list of available
protocols can be found at www.chemometec.com
2.1.3 The RUN button and LED indicator
On the front of the NucleoCounter® NC-202™ there is a RUN button. Activating this will run
the selected type of analysis on the instrument. In this way, it is easy to perform several
identical analyses in a routine manner.
2.1.4 Cassette fixture
The NucleoCounter® NC-202™ has a new cassette fixture design, allowing the Via2-
Cassette™to be inserted without any resistance. Executing a protocol will close the
cassette fixture by electromagnetic force, ensuring perfect alignment of the cassette
counting chamber with the focal plane.
Before closing the cassette fixture, a piston will press the Via2-Cassettes™plunger, driving
the cells through the staining channels and into the counting chamber.
These two mechanisms promote data consistency by using identical loading and focus
parameters between samples.
Refer to section “5.1 Cleaning” for guidelines on how to clean the fixture.
2.1.5 USB 3 connection
At the rear of the instrument, a USB data connection (rev 3.0 or higher), a DC power
connection and a DC fuse holder can be found. Please always take care of these areas and
keep them free of liquids and foreign objects.

2 Cell Counting with NucleoCounter® NC-202™
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2.2 The Via2-Cassette™
Figure 1: The Via2-Cassette™contains two fluorescent dyes, AO (Acridine Orange) and
DAPI (4',6-diamidino-2-phenylindole), deposited in the first part of the flow system
2.2.1 Loading the Via2-Cassette™ cassette
The Via2-Cassette™is loaded by pressing the white piston down while the cassette tip is
immersed in the cell sample (Figure 3). Pressing the piston creates a vacuum that draws
approximately 60 µl cell sample into the cassette flow system. The Via2-Cassette™is
designed for microcentrifuge tubes and accepts a minimum sample volume of 100 µl.
Figure 2: Loading the Via2-Cassette™
After loading the Via2-Cassette™, the sample buffer dissolves the fluorescent dyes, AO
(Acridine Orange) and DAPI (4',6-diamidino-2-phenylindole), deposited inside the flow
channels. The dyes instantly dissolve and stain the loaded cell sample.
After loading the Via2-Cassette™, it is inserted into the NucleoCounter® NC-202™
instrument. Pressing the RUN button will execute the cell counting protocol, that directs

2 Cell Counting with NucleoCounter® NC-202™
991-2020 Rev. 1.1 November 2019 7
the instrument to i) move the piston to drive the cell sample through the mixing channels
and into the measurement chamber; ii) fixate the cassette in the slot; and iii) record the
fluorescence images, analyze data and present the counting result.
Once analysis is complete, the sample and dyes are contained inside the cassette, which
can be safely discarded. This offers unique and safe sample handling and disposal.
The NucleoCounter® NC-202™ does not need cleaning or maintenance, since the cell
samples are not in direct contact with the instrument. The cell sample is contained within
the Via2-Cassette™and disposed of along with it. For regulated processes it is
recommended to perform periodic validation checks to document nominal performance of
the NucleoCounter® NC-202™ instrument and Via2-Cassette™.
2.2.2 Via2-Cassette™volume calibration dot code
During Via2-Cassette™production, the precise depth of the measurement chamber is
measured for every individual cassette, and a dot code is printed on the cassette. This dot
code is read by the NucleoCounter® NC-202™instrument and used to convert the absolute
cell number into a cell concentration.
2.2.3 Cassette handling
The image analysis method of the NC-View™offers considerable stability and can handle
minor scratches and fingerprints on the cassette windows. However, cell count quality is
best assured by keeping the windows of the cassette as clean as possible. The image
analysis will search for air bubbles, compensate the data, and flag the data point as
containing a bubble.
Be careful when attempting to wipe foreign objects off the cassette surface since the
plastic material of the cassette can be scratched. See how to place and remove the
cassette in section 2.2.1 “Declaration of Conformity”.

3 NucleoCounter® NC-202™ – Instrument Controls
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3NucleoCounter® NC-202™ – Instrument Controls
3.1 Interactive Controls
Figure 3: Front of the NucleoCounter® NC-202™
The NucleoCounter® NC-202™has two operational areas:
•Cassette fixture covered by a yellow lid
•RUN button and tri-color LED status indicator
The lid is secured to the NucleoCounter® NC-202™instrument with a magnet and can be
either opened or removed entirely during operation, exposing the cassette fixture.
The cassette is placed in the cassette fixture and locked in position by an electromagnet
when running a protocol. Fixing the cassette during acquisition ensures that the
measurement chamber and, in turn, the cells are always at the best focal setting.
The RUN button and LED indicator are placed to the right of the lid. The RUN button is
used to execute the protocol selected in the NC-View™software.
The LED indicator shows the instrument status:
•Green - the NucleoCounter® NC-202™is ready for the next operation
•Red - the NucleoCounter® NC-202™is busy or not connected
•Orange - the NucleoCounter® NC-202™has lost connection to NC-View™

3 NucleoCounter® NC-202™ – Instrument Controls
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3.2 NucleoCounter® NC-202™ Connections
Rear view of the instrument (Figure 5) features the DC power inlet, fuse holder (Figure 6)
and a USB 3.0 socket (Figure 7). The NucleoCounter® NC-202™is delivered with a
region-specific power cable to connect the external power supply to the main power
supply.
Figure 4: Rear view of the NucleoCounter® NC-202™
3.2.1 NC-202™power connection
The NucleoCounter® NC-202™ is powered by a 24VDC external power supply (Figure 6),
accepting between 100 - 240 V. It is not necessary to turn off the NucleoCounter®
NC-202™ between uses. When idle (Ready mode) the NucleoCounter® NC-202™ uses only
7 W.
Figure 5: DC power socket

3 NucleoCounter® NC-202™ – Instrument Controls
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3.2.2 USB connection and initialization
The NucleoCounter® NC-202™is connected using a USB 3.0 micro B –A or B –C cable. Only
cables provided by ChemoMetec should be used with the instrument.
Figure 6: USB 3.0 port
Launching the NC-View™software commences a short initialization sequence, which
includes an instrument communication check and positioning of the piston motor. After
initialization, the LED status indicator turns green and the NucleoCounter® NC-202™is
ready for operation. NC-View™software connection is indicated by the changing color of
the NC-View™ icon, and activation of the Run and Eject icons on the software
main window.

4 Operation of NC-View™ Software and NucleoCounter® NC-202™ Instrument
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4Operation of NC-View™ Software and
NucleoCounter® NC-202™Instrument
4.1 NC-View™ Software
The NC-View™ software is used for data acquisition, image analysis and display of results.
For further details regarding NC-View™, refer to the NC-View™Software User Guide (#991-
2022).
4.2 Protocol Execution
Once the NucleoCounter® NC-202™and the NC-View™software are connected, analysis
can begin. In the main window, the Run icon in the NC-View™ software or the RUN
button on the instrument can be used to execute the protocol of interest.
A standard operating procedure for NucleoCounter® NC-202™should include the following
steps:
1. Select protocol
2. Enter sample ID
3. Mix the cell sample by inverting sample tube a few times
4. Load the Via2-Cassette™with the cell sample
5. Place the cassette into the NC-202™cassette fixture and close the lid (fig. 8)
6. Press RUN
7. Review the image data
8. View the displayed results
After analysis, the cassette is removed from the fixture and disposed of as biological waste
according to national or regional regulations.
Figure 8: How to hold and place the Via2-Cassette™in the fixture

4 Operation of NC-View™ Software and NucleoCounter® NC-202™ Instrument
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4.2.1 Piston movements
When a cassette is analyzed, the piston motor will move the piston rod down through the
cassette cylinder. Hereby, the sample is transported to the cassette’s measurement
chamber while sensors monitor the sample flow. When the sample has been loaded into
the measurement chamber and reaches the end of the flow channel, the piston is stopped.
4.3 Protection of the Optical System
The lid covering the cassette fixture is designed to protect the analysis area from dust and
other potential contaminants and keep external light from interfering with fluorescent
image recording. The lid is mounted with a magnetic hinge.
The lid should always be closed to protect the instrument’s optical system, except when
inserting or removing a cassette, or cleaning the fixture.

5 Maintenance of NucleoCounter® NC-202™
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5Maintenance of NucleoCounter® NC-202™
5.1 Cleaning
5.1.1 Instrument cabinet
The NucleoCounter® NC-202™cabinet should be cleaned at regular intervals by gently
wiping the surface with a soft cloth and a mild detergent. Never use organic solvents or
aggressive detergents to clean the exterior of the NucleoCounter® NC-202™as this might
damage the surface.
5.1.2 Cassette fixture
When cleaning the cassette fixture, take great care to avoid introducing any liquid or dust
into the NucleoCounter® NC-202™. Any liquid entering the interior of the NucleoCounter®
NC-202™can damage the optical parts and compromise the quality of the cell count.
The black surface of the cassette fixture is anodized aluminum and can therefore be wiped
off with a clean, dry, and dust free cloth, reducing fiber contamination of the cassette
fixture.
5.1.3 Contamination of the optics
Foreign objects attached to the optical component can influence the collected image and
may be visible as faint objects in the image.
Presence of a contaminant (e.g. dust particles) will normally not influence the counting of
cells. The NucleoCounter® NC-202™will distinguish between cells and contaminants.
If contamination is visible in every image acquired, it indicates that the optical parts are
compromised. Compressed air (clean and dry) can be used to remove dust particles from
the optical system by blowing air into the cassettes fixture. Do not use compressed air
from a single use-can, since these contain chemicals that can reduce optical performance.
ChemoMetec’s support should be contacted if the contaminant cannot be removed with
compressed air.
5.1.4 Decontamination
If a complete decontamination of the NucleoCounter® NC-202™is required, please contact
ChemoMetec before taking action.