ChemoMetec NucleoCounter NC-250 User manual

NucleoCounter
NC-250
™
Quick Guide
Viability and Cell Count, Cell Health and Cell Cycle
in one instrument!
®

How to get started
Dear NucleoCounter Customer,
Thank you for purchasing the NucleoCounter®NC-250™which offers not only fast and precise
Viability and Cell Count Assays, but also fast and easy assaying of cell cycle profiles with top
class precision and the fastest apoptosis assay!
1
The NucleoCounter®NC-250™Concept
NucleoView™ Software
Included on a USB stick
for an unlimited number of
installations.
NC-Slides A8™and A2™
A8 slides for fast analysis and A2 slides
for high precision.
How to get started - 8 easy steps to install the NC-250™
Solution 18
For Viability and Cell Count
Assays.
 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 determination via the fastest
apoptosis assay
 Fast and easy operation
 Excellent reproducibility
 No calibration
 Maintenance and service
free
 21 CFR Part 11 ready
For detailed instructions, please read the manual.
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 installation of
the NucleoView™ NC-250 software.
Insert the USB stick in the PC and open the “Install_Guide.html” file for detailed
installation instructions.
Open the “Install NucleoView NC-250 X.X.X.X.exe” file (the Xs indicate the version
number e.g. 1.0.22.0). Do NOT open the .bin file.
Follow the instructions on the screen. After the software installation it will be
required to restart the PC.
Open the NucleoView™ NC-250 software by double clicking on the NC-250 icon on
the desktop to start the installation of the instrument.
Follow the on-screen instructions to install the instrument.
The NucleoCounter® NC-250™ is ready to use when the LED indicator light on the
instrument turns green it is ready to use.

2
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 counting.
Select the desired protocol from the menu:
for example, ‘Viability and Cell Count Assay’
Choose the media type (A8 or A2 slide)
Select the chamber(s) to alalyze
Optional: enter Sample ID and Operator Name
Mix the cell sample with the assay defined solution(s), load the slide and press
‘RUN’
TIP: Find the application note for the selected protocol by clicking the icon.

3
Optional: Visual Inspection
of Counting Gates
OPTION
(PREFERRED)
Click the green dot in the right-hand corner of the image window.
This activates the image overlay function indicating all the events in
the total cell count. The mouse scroll button allows the user to zoom
in at the cursor position. Cells will be framed by a pink square.
OPTION
(ADVANCED)
Right-click on the sample file name in the ’Data folders and
files’ window and select ’Show counting gates in Plot Manager’.
A new window will open displaying scatter plots and histograms of
event intensity and size for the appropriate channels (AO and DAPI).
It is important that the core of the population, usually seen as a coloured
region on the scatter plot, is included in the square counting gate.
Note it is not required that all events be included in the counting
gate. Events in the lower, left-hand corner of the scatter plot are often
smaller than cells and considered debris.

4
How to Perform a 2-Step
Cell Cycle Analysis
The NucleoCounter®NC-250™provides fast and easy assaying of cell cycle profiles with top
class precision.
Wash 5x105to 1x106cells for cells in suspension or adherent cells in a 6-well
plate with PBS
Load the slide and Press ‘RUN’
Add 20 μl Solution 12 to 980 μl Solution 10 and mix
Add the Solution 10 and 12 mixture to your adherent cells or cells in suspension
Incubate at 37°C for 5 minutes
Add same volume of Solution 11
Select the protocol from the menu: ‘2-Step Cell Cycle Assay’ and choose the
media type (A8 or A2 slide)
Add markers to the DNA content histogram to demarcate cells in the different
cells cycle stages.
Select the chambers to perform the assay on
Optional: enter Sample ID and Operator Name
Untreated HeLa cells Camptothecin treated HeLa cells Merged

5
How to Perform a Cell Health
Analysis
Select the protocol from the menu: ‘Vitality Assay’
Choose the media type (A8 or A2)
Select the chambers to perform the assay on
Optional: enter Sample ID and Operator Name
The NucleoCounter®NC-250™ provides cell health measurements by the world’s fastest
apoptosis assay. VitaBright-48™a component of Solution 6 instantly fluoresces when it
binds thiols inside cells. The level of thiols is a direct measurement of cell vitality and a
drop in the thiol level is concomitant with apoptosis.
Mix the cell sample with Solution 6, immediately load the slide
and press ‘RUN’
If required, the percentage of the different cell populations can
be investigated by adding a quadrant or markers to the scatter
plot or histogram, respectively
Untreated YAC-1 cells
Necrotic Cells
Camptothecin treated
YAC-1 cells
PI negative cells
with low vitality Healthy cells

6
Only if Required: Create your own version of
a Protocol with Adjusted Generic Gates
Perform the desired type of assay on a sample of the cells to be analysed.
In the Tools menu select Protocol Adaptation Wizard.
Follow the instructions in the Protocol Adaptation Wizard to create your own
adapted protocol.
The viability and cell count results will not be adjusted for the image file
used for the adaption of counting gates. New results can only be obtained
by running a new sample with the new gating protocol.
PDF reports
Right-click on file to create a PDF report
Select the parameters to be visible on the report, and the properties of the
parameters
Optional: preview your result
Save and/or print your report to the default printer

Additional Resources
ChemoMetec A/S
Gydevang 43
DK-3450 Allerod
Denmark
Phone[phone]
Fax [phone]
Mail [e-mail]
Web www.chemometec.com
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 assure their accuracy, ChemoMetec A/S assumes
no liability resulting from errors or omissions in these documents, or from the use of the information contained herein.
ChemoMetec A/S reserves the right to make changes in the product design without reservation and without notification to
its users.
Copyright Notices
Copyright © ChemoMetec A/S 2012. All rights reserved. No part of this publication and referred documents may be repro-
duced, stored in a retrieval system or transmitted in any form or by any means, electronic, mechanical, photocopying, record-
ing 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 A/S.
NC-Slides and NucleoView are trademarks of ChemoMetec A/S.
All other trademarks are the property of their respective owners.
Consumables/kits:
Item no. Description
942-0003 NC-Slides A8™
942-0001 NC-Slides A2™
910-3003 Solution 3
910-3006 Solution 6
910-3010 Solution 10
910-3011 Solution 11
910-3012 Solution 12
910-3017 Solution 17
910-3018 Solution 18
910-0003 Reagent A100
910-0002 Reagent B
912-0012 NucleoCounter®NC-250™IQ/OQ Kit
974-0004 NucleoCounter®NC-250™PQ Test Kit - Slides
Go to www.chemometec.com to find:
991-0253A ver. 01.2016
• Documentation
• SDS
• Application notes
• Certificates of analysis
• Videosetc.