NanoEnTek ADAM-MC User manual

A New Standard of
Automatic Cell Counter
ADAM
INSTRUCTION MANUAL
NESMU-AMC-001E (V.5.5)
A New Standard of
Automatic Cell Counter
ADAM-
MC
USER MANUAL
NESMU-AMC-001E (V.5.1)

2
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Table of Contents
Product Contents
Safety Information
Product Specifications
Description of ADAM-MC
3
4 ~ 6
7
8 ~ 9
Introduction
Overview
Basic principals of counting
10 ~ 11
10
11
Getting Started
Environmental requirements
Installation
Start-up screen
Error message during booting
Menu setting
System information
Chip selection
Counting method
12 ~ 18
12
13
14
15
16
16
17
18
General Operation
Introduction
Preparing cell
Counting cell
Operating ADAM-MC
Icon function
Result analysis
Error code
Maintenance and cleaning
19 ~ 24
19
19
19
20
21
22
23
24
Software Installation
Introduction
Getting started
Installation
Function guide
Function buttons
Data list
Information
Graph
Sample image of error message
25 ~ 32
25
25
26 ~ 27
28
29
30
30
31
32
Trouble shooting
Warranty
Product list
Purchaser notification
33
34
35
36

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ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Product Contents
Section 1
ADAM-MC The contents of the ADAM-MC are listed below:
Accuchip kit The contents of the ADAM-MC‟s Accuchip Kit are listed below:
Upon
receiving
the
instrument
•Examine the instrument carefully for any damage incurred during
transit.
•Ensure that all parts of the instrument including accessories listed
above are included with the product.
•Any damage claims must be filed with the carrier.
•The warranty does not cover in-transit damage.
•See the 13 page to install the instrument.
•Upon receipt, store Accustain solution at room temperature.
Item Quantity
Main device 1
User manual 1
USB cable 1
Installation CD 1
Key pad 1
Power cord 1
Fuse 2
External video monitor (Optional) 1
Item Accuchip2x Kit
(Cat. No: AD2K-200)
Accuchip4x Kit
(Cat. No: AD4K-200)
Accustain
Solution
(Cat. No:
ADR-1000)
Disposable
Chip 200pcs (2 channel) 200pcs (4 channel) N/A
Solution T 2 x 12.5 ml 2 x 12.5 ml 4 x 12.5 ml
Solution N 1 x 12.5 ml 1 x 12.5 ml 2 x 12.5 ml
Available
test Q‟ty
Min. 200 test/kit Min. 400 test/kit
Max. 400 test/kit
(Only total cell count)
Max. 800 test/kit
(Only total cell count)

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ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Safety
precautions
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
Always ensure that the power supply input voltage match the voltage
available in your location.
For operation environment, See page 12.
This machine is air-cooled so its surfaces become hot during operation.
When installing it, leave a spaces of more than 10 cm (4 inches) around it.
Never insert metallic objects into the air vents of the instrument as this
could result in electrical shock, personal injury and equipment damage.
Always set the main switch on the power supply unit to off before
connecting the power cord to the wall outlet.
Always ensure that the grounding terminal of the instrument and that of
the wall outlet are properly connected. The power cord should be
connected to a grounded, 3-conductor power outlet.
To avoid potential shock hazard, make sure that the power cord is properly
grounded.
Do not position the equipment so that it is difficult to operate the
disconnecting device.
Be sure to set the main switch to off, unplug the power cord and lock the
stage before moving.
If the instrument is broken or dropped, disconnect the cord and contact a
authorized service person. Do not disassemble the instrument.
Use only authorized accessories.
Use this equipment only as specified in this manual and as specified in any
documentation associated with its components. Any use of the equipment
in an unspecified manner is strongly discouraged and may result in
damage or injury as cautioned by signed warnings.
Section 3 Safety Information

5
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Safety
symbols
The symbols used in the ADAM-MC and the manual are explained below:
The Caution symbol denotes a risk of safety hazard.
ON (Power)
Protective earth (Ground)
The CE mark symbolizes that the product conforms to all applicable European
Community provisions for which this marking is required. Operation of the
ADAM-MC is subject to the conditions described in this manual.
The protection provided by the device may be impaired if the instrument is
used in a manner not specified by the manufacturer.
Caution, Biohazard
Protective measures must be used in dealing with biologically hazardous
materials such as carcinogenic reagents.
Disposal of your old appliance
1. When this crossed-out wheeled bin symbol is attached to a product it means
the product is covered by the European Directive 2012/19/EU.
2. All electrical and electronic products should be disposed of separately from
the municipal waste stream via designated collection facilities appointed by the
government or the local authorities.
3. The correct disposal of your old appliance will help prevent potential
negative consequences for the environment and human health.
4. For more detailed information about disposal of your old appliance, please
contact your city office, waste disposal service or visit our web-site,
www.nanoentek.com.
Safety Information,continued
Section 3

6
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Warnings Battery inside device
•Risk of explosion if battery is replaced by an incorrect type.
•This battery is not replaceable by a user.
•Refer to a qualified personnel.
Cover
•Do not remove a cover or dissemble a case.
•There is no adjustable components inside the instrument.
•If malfunction is found, refer to a service personnel.
Manual
•Do not attempt to service the equipment unless
this manual has been consulted and is understood.
•This manual is available in English only.
•Failure to heed this warning may result in injury to
service provider, operator from electric shock,
mechanical or other hazards.
Sample handling
•Wear gloves during sampling. User‟s sample may
have the infectious biohazardous substance.
Waste
•After using Accuchips, appropriately dispose it as
biohazardous waste.
•Do not reuse the Accuchips.
Operator
•Must have the general knowledge of cell counting
procedure and bio safety to handle the sample that
may have the infectious biohazardous substance.
Safety Information, continued

7
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Product Specifications
ADAM-MC Voltage AC 100 ~ 240 V,
50 ~ 60 Hz
Current max. 1.8 A, max 100 W
Fuse F3.15AL250V
Objective lens 4 X
LED Green LED
CCD camera B/W CCD
Filter Excitation filter
Dichroic filter
Emission filter
Weight 9Kg
Size (W×L×H) 220 ×375 ×250 mm
Degree of
protection
IPX0
AccuChip AccuChip 2x AccuChip 4x
Measuring range 5ⅹ104~ 4ⅹ106cells/mL
Analysis time 45 sec. ~ 2 min./test
Loading sample
vol. per test
20 µ L/test (AccuChip 2X)
12 µ L/test (AccuChip 4X)
Measuring
sample vol. per
test
8.5 µ L/test (AccuChip 2X)
3 µ L/test (AccuChip 4X)
AccuStain
solution T,N
PI (Propidium Iodide) staining of
total cells (T) and non-viable cells (N).
Accessories
Power cord 1.5 m
Fuse 250 VAC, 3 A;
F3.15AL250V
Keypad USB Type / 1.3 m
Section 1

8
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Front view
of ADAM-MC
The ADAM-MC, an automated cell counter, is a benchtop automated
cell counter that performs cell count and viability measurements
using Accustain solution.
The front view showing various parts of the ADAM-MC is shown below:
•Control buttons:
Eject Ejects the chip holder from the ADAM-MC.
Start Performs all procedures of automatic counting.
Lock Protects the alignment of stage from external shock
when the ADAM-MC is moved to the other places.
☞It is strongly recommended to lock ADAM-MC before turning it off.
•Door Chip holder comes out here.
•LCD Displays the process and the result.
•keypad Inputs the sample number less than 3 characters
and press „Enter‟ button.
Door
LCD
Key Pad
Control Button
Section 1 Description of ADAM-MC

9
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Rear view
of ADAM-MC
The rear view showing various parts of the ADAM-MC.
Fan Cooling fan
Power switch Main power on/off switch
Power plug ADAM-MC power cord connection plug
USB port Connect to computer with USB Cable
RS232C port Not connect (Port for only QC and Service)
Key pad port Keypad connection port
Video Port External video monitor port
Section 1 Description of ADAM-MC,continued
Power
Switch
Power
Plug
Video
Port
USB
Port
RS232C
Port
Key Pad
Port
Fan

10
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Overview Until now, cell counting and viability measurement for many types of cells have
been performed manually using hemocytometer with Trypan Blue exclusion
method, which is to distinguish viable cells from non-viable cells. One drawback
of this method, however, is the propensity for the staining of artifacts; another
drawback is that the naked eye can only differentiate between cells in a limited
concentration range in the hemocytometer chamber. This combined with the
potential problem of cell aggregation and limited sample volume leads to the
common variation of counts normally associated with this method.
To address these problems, NanoEnTek has developed the ADAM-MC, which is
based on a fluorescent microscopy technique for counting cells. The ADAM-MC
utilizes sensitive fluorescence dye staining, LED optics and CCD detection
technologies to make the cell analysis more accurate and reliable.
To count cells using ADAM-MC, the cells are mixed with a Propidium Iodide (PI)
stain and directly pipetted on to a disposable plastic chip. The chip is
then loaded onto a precision stage. An ADAM-MC system is automatically
focused onto the chip and cells that have been stained are recorded by
a sensitive CCD camera. The image results are automatically processed
generating the cell count which is displayed on the front of the instrument.
Simple. Fast. Accurate. Reliable.
Introduction

11
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Basic
principals of
counting
ADAM-MC is based on staining mammalian cell DNA with a fluorescent dye,
Propidium Iodide (PI). PI does not enter cells with intact cell membranes
or active metabolism. In contrast, cells with damaged membranes or with
inactive metabolism are unable to prevent PI entering the cell. As a result,
the nuclei of non-viable cells will only be stained. The ADAM-MC provides two
kinds of staining solutions. AccuStain Solution T for the total cell counting is
composed of the fluorescent dye (PI) and lysis solution. AccuStain Solution N for
the non-viable cell counting is composed of the fluorescent dye and PBS.
In order to measure the total concentration of cells, the plasma membranes of all
the cells must be disrupted to stain all the Nuclei with PI. The process of
disrupting and staining is achieved by treatment with AccuStain Solution T.
In the second solution, live cells remain intact and are not stained.
Only the non-viable cells are stained and detected. After treatment, the prepared
cells will be loaded into the chip. The viability will be automatically calculated in
the ADAM-MC software after each measurement of the total cells and the non-
viable cells.
Introduction,continued
A: Total cell
B: Non-viable cell

12
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Environmental
requirements
To obtain the best results,
install the ADAM-MC in a location following conditions:
1. Room temperature between 20 and 35 °C.
•Not recommended for cold room use (≤ 4 °C).
•At low temperature (≤10 °C), warm up the ADAM-MC for 10 min.
2. Not exposed to direct sun light.
3. Not subject to direct or continuous vibration..
4. Not subject to intense magnetic or electromagnetic fields.
5. Relative humidity between 0–95 %.
6. Area free from corrosive gases or other corrosive substances.
7. Area with very little dust or other airborne particles.
8. Allow a 10 cm minimum space around the instrument
for proper air flow.
9. Not allow to put heavy material on top of ADAM-MC.
Getting Started

13
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Installation 1. Remove all components of ADAM-
MC from their box.
2. Place the instrument in a flat
level and dry surface.
3 Plug to power cord into the
electrical outlet.
•Be sure to use only the
power cord supplied with
your instrument.
•Powering the instrument with
an unapproved power cord
may damaged the instrument.
4. Plug to keypad.
5. turn on the power switch.
•Make sure that the main
power switch is In the “Ⅰ”
(ON) position.
Getting Started,continued
Section 3

14
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Start-Up
screen
1. System booting.
2. System will go through self
diagnostic tests.
•If you get an error message,
please contact your local
distributor or
[e-mail].
3. The home screens will be
displayed as the image, no errors
are detected.
Section 3 Getting Started, continued

15
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Error
message
during
booting
1.
2.
3.
It appears when booting not
working properly.
Turn off main power and
restart device.
If this message still appears after
restart, contact your local
distributor or
[e-mail].
1.
2.
3.
It appears when a chip is inserted
during Start up.
Remove the chip from a device,
and Do not turn on the device
with a chip.
If this message continues to
display when no chip is inserted,
contact your local distributor or
[e-mail].
1.
2.
It appears when turning off
without locking device.
Push the lock button before turn
off the device. If this message still
appears after
restart, contact your local
distributor or
[e-mail].
Getting Started,continued
Section 3

16
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Menu
setting
1.
2.
You can set the menu as you
press the „*‟ button on the keypad
from the screen for inputting
sample numbers.
You can select the number from
the Menu.
System
information 1.
2.
3.
Check the device versions and
date which have been installed in
the device.
Select number 4 key from the
MENU, and Press the „Enter‟ key.
The screen will return to the Menu
screen automatically.
Section 3 Getting Started,continued
[System Info.]
1. Version : MC V1.21
2. Date : Sep 23, 11
CORE1 : 1.87
CORE2 : 1.52
CORE3 : 1.53

17
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Chip
selection
1. Press „1‟ for chip selection.
You can select the number from
the Menu.
2.
3.
User can select the 2 kinds of chip
type.
•„1‟ : Two channel chip
(AccuChip2X)
•„2‟ : Four channel chip
(AccuChip4X).
Press „1‟ or „2‟, and Enter key.
4. After pressing the „Enter‟ key,
the screen will return to the
Menu screen automatically.
Getting Started,continued
Section 3

18
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Getting Started,continued
Section 3
Counting
method
1. Press „2‟ for counting method.
You can change the capture frame
mode.
2. You can select the 60 or 30
frames in 2CH mode and 22 or 11
frames in 4CH mode for counting
as well.
2CH
High Precision: 60 frames capture
High Speed : 30 frames capture
4CH
High Precision: 22 frames capture
High Speed : 11 frames capture
3.
4.
Press „1‟ or „2‟ and „Enter‟ key.
After pressing the „Enter‟ key,
the screen will return to the Menu
screen automatically.

19
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
Non-viable
Total
Introduction Instruction is provided in this section for preparing the
cell sample with Accustain solution for use with disposable
Accuchip for automated cell count using the ADAM-MC.
Preparing
cell
1) Cultivate the required number of cells.
2) Add an appropriate volume of growth media or PBS to dilute
to a final concentration of 5ⅹ104cells/ml to 4ⅹ106cells/ml.
Note ☞ Concentration out of this range will result in errors.
Refer to page 23 for more information about errors.
3) Thoroughly mix the cell pellet by vortexing.
4) Check visually if any cell clumps or agglomerates are remained.
Counting
cell
Accuchip 2x
Total Cell Non-viable Cell
1) Add 50 µ L of your sample to 50 µ L
supplied Accustain solution T.
2) Mix gently by pipetting up and down.
3) Load 20 µ L sample mixture to the
Accuchip on T channel.
1) Add 50 µ L of your sample to 50 µ L
supplied Accustain solution N.
2) Mix gently by pipetting up and down.
3) Load 20 µ L sample mixture to the
Accuchip on N channel.
Accuchip 4x
Total Cell Non-viable Cell
1) Add 50 µ L of your sample to 50 µ L
supplied Accustain solution T.
2) Mix gently by pipetting up and
down.
3) Load 12 µ L sample mixture to the
Accuchip on T1 or T2 channel.
1) Add 50 µ L of your sample to 50 µ L
supplied Accustain solution N.
2) Mix gently by pipetting up and down.
3) Load 12 µ L sample mixture to the
Accuchip on N1 or N2 channel.
Section 4
Note ☞ When you load of the sample mixture to the Accuchip, please be careful not to make bubbles.
Non-viable
Total
Non-viable
Total
General Operation

20
ADAM-MC (Automated Cell Counter) ⓒ2008 NanoEnTek Inc.
General Operation,continued
Section 4
Operating
ADAM-MC
1. Press “ Eject(▲)” button on
the main device to eject the
chip holder.
2. Insert the Chip loaded with the
sample onto the chip holder.
3. Press the “Start(▶) ” button on
the main device.
Note ☞ Automatic Focus will be carried out at the first time the device is booted.
Once ADAM-MC has done the Auto Focus process and on the following time,
focusing process will be skipped.
4. The instrument takes
approximately 90 seconds to
count sample.
5. After calculating the cell number,
the chip will be ejected
automatically.
Then chip can be removed.
6. The calculated cell number per
1ml will be displayed on the
screen automatically.
7. For another experiment, repeat the process from steps 1 ~ 5.
ID1 : 123 ID2 : 456
T1=1.10E6 N1=5.50E5
T2=2.20E6 N2=5.50E5
T1-N1=5.50E5 V1= 50%
T2-N2=1.65E6 V2= 75%