ChemoMetec NucleoCounter NC-250 User manual

Quick Guide
Viability and Cell Count, Vitality and
Cell Cycle Assay in one instrument!
NucleoCounter®NC-™

How to get started
Dear NucleoCounter®Customer,
Thank you for purchasing the NucleoCounter® NC-250™ which oers not only fast and precise
Viability and Cell Count Assays, but also fast and easy assaying of cell cycle proles with top class
precision and the fastest apoptosis assay!
How to get started - 8 easy steps to install the NC-™
Unpack the NC-250™ instrument and plug it in the main outlet. Do NOT connect the USB
cable to the PC.
Make sure that there are full administrator rights on the PC during the installaon of the
NucleoView™ NC-250™ soware.
Insert the USB sck in the PC and open the “Install_Guide.html” le for detailed installaon
instrucons.
Open the “Install NucleoView NC-250 X.X.X.X.exe” le (the Xs indicate the version number
e.g. 1.0.22.0). Do NOT open the .bin le.
Follow the instrucons on the screen. Aer the soware installaon it will be required to
restart the PC.
Aer the restart the NucleoView™ NC-250™ soware will automacally open and nish the
installaon.
Follow the on-screen instrucons to install the instrument.
The NucleoCounter® NC-250™ is ready to use when the LED indicator light on the instrument
turns green.
For detailed instrucons, please read the manual.
NucleoView™ Soware
Included on a USB sck for an
unlimited number of installaons.
Soluon 18
For Viability and Cell
Count Assays.
NC-Slides A8™ and A2™
A8-Slides™ for fast analysis and
A2-Slides™ for high precision.
The NucleoCounter®NC-™ Concept
 8 viability and cell counts in less than 3 minutes
- Total cell count
- Viability
- Cell diameter
- % of cells in clumps
 Up to 8 advanced analyses in one run
- Fast and easy - High precision cell cycle analysis
- Cell health determinaon via the fastest apoptosis assay
 Fast and easy operaon
 Excellent reproducibility
 No calibraon
 Maintenance and service free
 21 CFR Part 11 ready
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How to perform Viability
and Cell Count Analysis
Fast and Precise Cell Count and Viability Assays!
The NucleoCounter® NC-250™ from ChemoMetec is a step forward for automated cell counng.
Select the desired protocol from the menu: for example, ‘Viability and Cell Count Assay’
Choose the media type (NC-Slides A8™ or A2™)
Select the chamber(s) to analyse
Oponal: Enter Sample ID and Operator Name
Mix the cell sample with the assay dened soluon(s), load the NC-slide and press ‘RUN’
TIP: Find the applicaon note for the selected protocol by clicking the icon.
 Fast and easy operaon
 Excellent reproducibility
 No calibraon
 Maintenance and service free
 21 CFR Part 11 ready
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Optional: Visual Inspection
of Counting Gates
OPTION 1 (PREFERRED)
Click the green dot in the right-hand corner of the image window.
This acvates the image overlay funcon indicang all the events in the total cell count. The mouse
scroll buon allows the user to zoom in at the cursor posion. Cells will be framed by a pink square.
OPTION 2 (ADVANCED)
Right-click on the sample le name in the ‘Data folders and les’
window and select ‘Show Raw Data’.
A new window will open displaying scaer plots and histograms
of event intensity and area for the appropriate channels (Acridine
Orange and DAPI).
It is important that the centre of the populaon most usually
seen as a colored region on the scaer plot, is included in the
counng gate.
To check disnct cell populaons, create a new polygon around
the parcular cell populaon.
Right-click inside of the newly formed polygon and select
‘Add Cells Inside Gate to Image Overlay’. This acvates the
image overlay funcon indicang all the events visually to
determine the validity of their inclusion or exclusion from the
nal counng result.
Delete the polygon and the image overlay by right-click and
selecng ‘Delete Image Overlay’.
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How to Perform a -Step
Cell Cycle Assay
The NucleoCounter® NC-250™ provides fast and easy determinaon of cell cycle proles with top
class precision.
Add 20 μl Soluon 12 to 980 μl Soluon 10 and mix
Make a cell pellet of 5x105 to 1x106cells
Add 250 µl of the Soluon 10 and Soluon 12 mixture to your adherent cells or cells in
suspension and mix
Incubate at 37°C for 5 minutes
Add 250 µl of Soluon 11 and mix
Select the protocol from the menu: ‘2-Step Cell Cycle Assay’ and choose the media type
(NC-Slide A8™ or NC-Slide A2™)
Select the chambers to perform the assay on
Oponal: Enter Sample ID and Operator Name
Load the NC-Slide A8™ or NC-Slide A2™ and Press ‘RUN’
Add markers to the DNA content histogram to disnguish between the cells in the dierent
cells cycle stages.
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Untreated HeLa cells Camptothecin treated
HeLa cells
Merged

How to Perform a Vitality Assay
The NucleoCounter® NC-250™ provides the world’s fastest Vitality assay. Vitabright48™ is a
component of Soluon 6instantly uoresces when it binds thiols inside cells. The level of thiols is a
direct measurement of cell vitality.
Select the protocol from the menu: ‘Vitality Assay’
Choose the media type (NC-Slide A8™ or NC-Slide A2™)
Select the chambers to perform the assay on
Oponal: Enter Sample ID and Operator Name
Mix the cell sample with Soluon 6(in a rao 19:1), immediately load the NC-Slide A8™ or
NC-Slide A2™ and press ‘RUN’
If required, the percentage of the dierent cell populaons can be invesgated by adding a
quadrant or markers to the scaer plot or histogram, respecvely
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Perform the desired type of assay on a sample of the cells to be analyzed.
In the Tools menu select Protocol Adaptaon Wizard or right-click on the desired le and
select ‘Start Protocol Adapon Wizard’.
Follow the instrucons in the Protocol Adaptaon Wizard to create your own adapted protocol.
Note: The viability and cell count results will not be adjusted for the image le used for the
adapon of counng gates. New results can only be obtained by running a new sample with the
new gang protocol.
Only if Required: Create your own version of a
protecol with Adjusted Generic Gates
PDF Reports
Right-click on le to create a PDF report
Select the parameters to be visible on the report, and the properes of the parameters
Oponal: Preview your result
Save and/or print your report to the default printer
Tip: Batch exports can be done from the NucleoView™ File Browser.
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Disclaimer Noces
The material in this document and referred documents is for informaon only and is subject to change without noce. While reasonable
eorts have been made in preparaon of these documents to assure their accuracy, ChemoMetec A/S assumes no liability resulng from
errors or omissions in these documents, or from the use of the informaon contained herein.
ChemoMetec A/S reserves the right to make changes in the product design without reservaon and without nocaon to its users.
Copyright Noces
Copyright © ChemoMetec A/S 2017. All rights reserved. No part of this publicaon, soware and referred documents may be
reproduced, stored in a retrieval system, or transmied in any form or by any means, electronic, mechanical, photocopying, recording or
otherwise, without the prior wrien consent of ChemoMetec A/S, Gydevang 43, DK-3450 Allerød, Denmark.
ChemoMetec© and NucleoCounter© are registered trademarks owned by ChemoMetec A/S.
NC-SlidesTM and NucleoViewTM are trademarks of ChemoMetec A/S.
All other trademarks are the property of their respecve owners.
Consumables/kits:
Item no. Descripon
942-0003 NC-Slides A8™
942-0001 NC-Slides A2™
910-3003 Soluon 3
910-3006 Soluon 6
910-3010 Soluon 10
910-3011 Soluon 11
910-3012 Soluon 12
910-3017 Soluon 17
910-3018 Soluon 18
910-0003 Reagent A100
910-0002 Reagent B
912-0012 NucleoCounter® NC-250™ IQ/OQ Kit
974-0004 PQ Package for NucleoCounter® NC-250™
• Documentaon
• Safety Data Sheet
• Applicaon notes
• Cercates of analysis
• Videos etc.
Additional Resources
Go to www.chemometec.com to find:
ChemoMetec A/S
Gydevang 43, DK-3450 Allerød - Denmark
Phone
Fax
Mail
Web
([phone]
([phone]
[e-mail]
www.chemometec.com
991-0253B ver. 01-2017