Olympus CKX-CCSW User manual

INSTRUCTIONS
NOTE:
This instruction manual is for the Olympus confluency checker CKX-CCSW.
To ensure the safety, obtain optimum performance and to familiarize yourself fully with the use of this
software, we recommend that you study this manual thoroughly before using the software, and always
keep this manual at hand when using the software.
Keep this instruction manual in a safe place.
CKX-CCSW
Confluency checker
Software for research/education
This software is designed to use in research and or education fields. Optical Microscope Accessory

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Introduction....................................................................................................................................................... 1
Handling Precautions .............................................................................................................................. 2
Cautions in use............................................................................................................................................................... 2
System compatibility.................................................................................................................................................. 2
PC and software ............................................................................................................................................................ 3
1 System diagram....................................................................................................................................... 4
2 Installation..................................................................................................................................................... 5
2-1 Installation................................................................................................................................................................ 5
2-2 Activation .................................................................................................................................................................. 8
2-3 Uninstallation ........................................................................................................................................................ 8
3 Basic operations..................................................................................................................................... 9
3-1 Initial Settings......................................................................................................................................................... 9
1Starting the software ..................................................................................................................................................................9
2Exiting the software..................................................................................................................................................................10
3Initial settings................................................................................................................................................................................. 11
3-2 Measuring while acquiring the image with DP22/DP27..............................................13
1
Measuring the cell count in the culture vessel while acquiring the image with DP22/DP27 ...... 13
2
Measuring the confluency in the culture vessel while acquiring the image with DP22/DP27 ... 17
3-3 Measuring from the acquired image............................................................................................ 21
1Measuring the cell count from the acquired image....................................................................................21
2Measuring the confluency from the acquired image.................................................................................24
3-4 Measuring while acquiring the image with cameras other than DP22/DP27 ......28
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Measuring the cell count in the culture vessel while acquiring the image with cameras other than DP22/DP27 ....28
2
Measuring the confluency in the culture vessel while acquiring the image with cameras other than DP22/DP27 ..32
3-5 Measuring the cell count and the confluency at the same time.........................36
3-6 Re-using the measurement settings ............................................................................................36
4 Camera connection (when using DP22/DP27)........................................................37
Contents

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CKX-CCSW
5 Screen flow...............................................................................................................................................38
5-1 [Main] screen......................................................................................................................................................38
5-2 Displaying the submenu..........................................................................................................................39
5-3 Displaying the [Setting] screen...........................................................................................................39
6 Various settings.....................................................................................................................................40
6-1 [Main] screen......................................................................................................................................................40
1Setting the source.....................................................................................................................................................................40
6-2 Submenu ............................................................................................................................................................... 41
1Setting the area of culture vessel................................................................................................................................41
2Setting the objective magnification ...........................................................................................................................41
3Setting the adapter magnification...............................................................................................................................42
4Setting the size of image sensor ................................................................................................................................42
5Setting the number of images........................................................................................................................................43
6-3 [Setting] screen.................................................................................................................................................43
1Setting the file format used for saving results..................................................................................................44
2Setting the count color..........................................................................................................................................................45
3Setting the confluency color ............................................................................................................................................46
4Displaying the focus indicator........................................................................................................................................47
5Setting the working folder..................................................................................................................................................48
6Setting the file name...............................................................................................................................................................48
7Setting the language...............................................................................................................................................................49
8Setting the DP image format............................................................................................................................................49
9Setting the calibration of DP22/DP27......................................................................................................................50
10 Displaying About.........................................................................................................................................................................50
6-4 [Property] screen ..............................................................................................................................................51
6-5 Saving the settings.......................................................................................................................................52
6-6 Loading the settings....................................................................................................................................52

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7 Effective measurement of cell count/confluency...................................................53
7-1 Zoom area ............................................................................................................................................................53
7-2 Using the cell scale bar effectively.................................................................................................53
1Setting the cell scale bar ....................................................................................................................................................54
8 Specifications..........................................................................................................................................55
9 Troubleshooting....................................................................................................................................56

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CKX-CCSW
Contents in this instruction manual
This instruction manual covers only the contents concerning the confluency checker CKX-CCSW. For procedures to
use the microscope digital camera DP22/DP27 or other cameras, refer to the instruction manual of the respective
camera.
Trademark
Microsoft, Excel, Internet Explorer and Windows are registered trademarks or trademarks of U.S. Microsoft
Corporation in the United States and other countries.
Intel and Intel Core are trademarks of Intel Corporation in the United States and/or other countries.
Company names and/or product names described in this manual are trademarks or registered trademarks of the
respective companies.
Software license agreement
If you use this product, you shall be deemed to have consented to the terms and conditions in “OLYMPUS END-
USER LICENSE AGREEMENT”.
"OLYMPUS END-USER LICENSE AGREEMENT" is included in the product. Please read it carefully before using the
product.
Introduction
The confluency checker CKX-CCSW is a product to measure the cell count and the confluency from the image where
the culture cells are acquired with the Olympus microscope digital camera DP22/DP27 or other cameras.

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Handling Precautions
Cautions in use
System compatibility
Image data
1. The recorded image data may be lost (destroyed) in the following cases. Olympus is not liable for the loss
(destruction) of the recorded data.
• When the user or a third party carried out a repair to the product.
• When the system is terminated or the power cord is unplugged while PC is recording or deleting (initializing).
• When the cable is disconnected while acquiring still images or movies.
• When the data have been kept saved in the recording media such as USB memory beyond its data storage
life (approx. from one to several years).
• When the product is damaged.
2. If following phenomena occur even though the camera is working properly, images cannot be restored and the
damages are not compensated.
• The image is abnormal.
• File properties such as file names, file date, etc. are abnormal.
• The image was lost.
} In general, as there is a data storage lifetime in the recording media, such as USB memory, HDD, CD-R, DVD-R, etc.,
the saved files may be lost after several years.
} The image data may be lost (corrupted) unexpectedly, so make frequent backups of the data you acquired.
OLYMPUS shall have no liability for any damage (including compensation for the corrupted image data) from the
use or incapable use of this product.
} Make sure that there is no problem in acquiring the specimen images before acquisition. Even though the problem
may occur by the acquired images, Olympus is not liable for these problems.
Connection to network
If you use this product connected to the network, the product may not work properly due to following problems.
1. Infection to the computer virus through network
2. Malfunction caused by applying OS service pack, security patch, or updating third-party software including runtime
libraries
3. The acquired images are missed or the acquisition process is delayed or stopped due to the overloaded system
which is caused by running the antivirus software, etc.
Restrictions in use
} Please also confirm the restrictions described in the instruction manuals of the digital camera DP22/DP27 or other
cameras you will use.

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CKX-CCSW
PC and software
Recommended configurations
Common in Desktop PC / Laptop PC / Tablet PC
No. Item Operating Environments
1 CPU Intel®Atom, Intel®Core.i3, Intel®Core.i5, Intel®Core.i7, Intel®Xeon, (CPU with the processing
capability of Intel®Core i3 or more is recommended.)
2 RAM 4GB
3 HDD 1 GB or more hard disk space for installation
4 Display Desktop PC / Laptop PC:
Resolution 1280 x 1024 (at least 1024 x 768), graphic card available for 32 bit color.
Tablet PC:
10.1 inch 1920 x 1200 (at least 8.3 inch)
5 Communication
port
USB3.0 port (when using DP22/DP27)
6 OS Microsoft®Windows®8.1 Professional (32bit/64bit)
Microsoft®Windows®8 Professional (32bit/64bit)
Microsoft®Windows®7 Ultimate/Professional (32bit/64bit) SP1
7 Language English, Japanese, Simplified Chinese
Even though the above mentioned recommended operating environments are satisfied, it does not necessarily
guarantee the functions of combination use with all PCs commercially available. Refer to Olympus website to check the
PCs whose performances in combination use with this system were confirmed.
PC
1. The PC data may be lost (corrupted) unexpectedly, so make frequent backups of the data you acquired. Olympus
is not liable for any damage due to the use or disability in use of this product, including compensation for lost data.
2. When the free space in the hard disk drive (HDD) is reduced, the data processing speed may be extremely slow or
errors may occur frequently. To prevent this, delete unnecessary data files frequently.

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1System diagram
The system configuration for using this product is shown below.
} Consult Olympus for the compatible microscope and camera adapter.
* The camera head may not work properly in combination with some PCs. Refer to Olympus website to check the
PCs whose performances in combination use with this system were confirmed.
** Do not use the interface cable for other use.
DP22-CU/DP27-CU
Interface cable **
PC *
(Desktop PC / Laptop PC /
Tablet PC)
CKX-CCSW
(Download from Olympus
web site.)
Microscope
・ CKX53 system, etc.
Camera
・ DP22-CU *
・ DP27-CU *
(Hereinafter referred to as
DP22/DP27)
Camera adapter
・ U-TV0.5XC
・ U-TV1X
・ U-CMAD
This product
Camera head

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CKX-CCSW
2Installation
2-1 Installation
Before installing software
1. Close all running applications before installing software.
2. Software cannot be installed unless the user account logging on the Windows is registered as a “Administrator”. If it
is registered as a “User”, change it to a “Administrator”.
(For the change of user account, refer to the Help of Windows.)
The procedures to install this product is described below.
Open (decompress) the file downloaded from the Olympus web site on the desktop screen of PC where this product
will be installed.
Click "Setup.exe".
Select the language to use.
Select the language from followings.
· English
· Chinese (simplified characters)
· Japanese
The procedures described below are for the case when [Japanese] is selected.
Click [Next].
Select [Yes, I accept all terms of the License Agreement.] and click [Next].
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Input [User name] and [Company].
These fields may have already been filled.
Select the folder to install the software and click [Next].
The following folder is specified as the initial setting.
32-bit version C:\Program Files\Olympus\Micro\CKX-CCSW
64-bit version C:\Program Files (x86)\Olympus\Micro\CKX-CCSW
Click [Install].
The installation starts. During the installation process, if "Would you like to install this device software?] regarding the
installation of the USB driver of DP22/DP27 appears, click [Install].
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CKX-CCSW
Click [Finish].
This is the end of the installation.
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2-2 Activation
2-3 Uninstallation
If passing 30 days of more after the date of installation, you must activate this product using the serial code. Without
activation, you will not be able to use this software.
The procedures to activate this product is described below.
The procedures to uninstall this product is described below.
If "Setup.exe" is deleted during installation, use Windows OS functions to uninstall the product.
TIP
Click on the desktop.
This product starts and the screen to input the license key appears.
Input the serial number in the [Serial No.] field a,
and input the license key in the [License Key] field b.
Click [Register] c.
Clicking [Finish] will close this screen without activation.
Within 30 days after the date of installation, the above screen
appears when exiting the software.
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TIP
Click "setup.exe".
Click [Next].
Select [Delete] and click [Next].
The uninstallation starts.
Click [Finish].
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b c
a

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CKX-CCSW
This section describes the basic operating procedures of this product. After initial settings are completed, you can
measure the cell count and the confluency according to the procedures described below.
· Measure the cell count/confluency in the culture vessel while acquiring the image using the camera (DP22/DP27, etc.).
· Measure the cell count/confluency in the culture vessel from the acquired images.
3Basic operations
3-1 Initial Settings
1Starting the software
Start this product according to the procedures described below.
· Windows 7
Click on the desktop screen.
Or click in the order of [Start] button - [All programs] - [Olympus] - [CKX-CCSW].
If the error or the warning screen appears after the software starts, click [OK].
· Windows 8/8.1
Display the desktop screen and click .
Or on the start screen, click [CKX-CCSW.exe] in [All applications].
If the error or the warning screen appears after the software starts, click [OK].
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Item Description
aControl area
Area to perform main operations of this product You can select from three
types of screens: [Main] / [Setting] / [Property] by the tab in the upper part
of the area.
bZoom area Area to zoom in/out the image in the preview area
cPreview area Area to display the image to be measured
dThumbnail area * Area to display the image in the specified folder by thumbnail and to
select the image to be measured
eFocus indicator * Displays the contrast level of the live image as a focusing guide when
acquiring with DP22/DP27.
fCell scale bar Scale to check whether the cell size is measurable or not
gButton to minimize Minimizes the screen.
hButton to close Closes CKX-CCSW
* These fields may not be displayed depending on the camera to be used and software settings.
2Exiting the software
Exit this product according to the procedures described below.
Click in the title bar.
Click [OK].
If the software has not been activated, the activation screen appears.
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e
b
c
f
a
d
gh
Entire view
When you start this product, the following screen appears first.

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CKX-CCSW
3Initial settings
This section describes the initial setting procedures of this product. This operation is necessary when you start this
product for the first time. After the second time, the product starts in the status reflecting the last settings when exiting the
software.
When you start this product, the message "Unable to display due to no recorded picture. Set the folder containing the
acquired image." may appear, but it is not a malfunction.
(Measuring while acquiring the image with the camera) Confirm that
powers of PC and the camera are turned OFF and connect PC to
the camera.
For the connection procedures when using DP22/DP27, see page
37. When using other cameras, refer to the instruction manual of the
respective camera.
(Measuring while acquiring the image with the camera) Confirm that
the light path selector of the microscope is set to the camera side.
For procedures to operate the light path selector of the microscope,
refer to the instruction manual of the microscope in use.
Start this product according to the procedures described on page 9.
The [Main] screen appears in the control area.
Click [Setting] a.
The [Setting] screen appears.
Click [General] b.
Click c.
The screen to select the folder appears.
Select the folder to use as a working folder and click [Select Folder].
The selected folder is used for saving the image acquired
with this product or for selecting the image to be used for
measurement. Images in the folder are displayed in the
thumbnail area of this product.
(These images are not displayed when measuring while
acquiring the image with DP22/DP27.)
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b a c

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Click hin [Size of Image Sensor] to display the items, and
select the size of the image sensor of the camera connected to the
microscope.
If DP22/DP27 is connected, settings are not necessary.
Items to be displayed are described below.
If [Custom] is selected, input the numerical value in the ifield.
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This is the end of initial settings.
No. Magnification
1 X0.5 (Initial setting)
2 X1.0
No. Sensor type [mm] (Horizontal) [mm] (Vertical)
1 DP22 – –
2 DP27 – –
3 4/3 17.3 13.0
4 Custom Direct input Direct input
Click [Main] d.
The [Main] screen appears.
In [Source], click eand select the source.
Select the source from followings.
· [DP]: Appears when DP22/DP27 is connected. (Keep this status
when measuring while acquiring the image with DP22/
DP27.)
· [Folder]: Appears when DP22/DP27 is not connected. (Keep
this status when measuring while acquiring the image
with other cameras.)
· [Select Image]: Select this field when measuring from the
acquired image.
If the submenu is not opened, click [Configuration] fto open the
submenu.
Click gin [Adapter Magnification] to display the items, and select
the camera adapter connected to the microscope.
Items to be displayed are described below.
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d f e
g
h i

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CKX-CCSW
3-2 Measuring while acquiring the image with DP22/DP27
You can measure the cell count and the confluency in the culture vessel while acquiring the image with DP22/DP27.
Start this product according to the procedures described on page 9.
The [Main] screen appears in the control area.
Click ain [Source] and select [DP].
Place the culture vessel of the specimen to be measured on the stage
of the microscope.
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3
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Measuring the cell count in the culture vessel while acquiring the image with DP22/DP27
While viewing the live image of the camera displayed in the preview
area, rotate the focusing knob of the microscope to bring the cell in
the culture vessel into focus. At this time, the blue bar of the focus
indicator must be 80% or more of the entire length of the focus indicator.
(If the blue bar is not 80% or more of the entire length, refer to the
instruction manual of the microscope and set each units of the
microscope, adjust the brightness, clean lenses, etc. properly.)
4
While viewing the live image of the camera displayed in the preview
area, rotate the nosepiece of the microscope and select the objective
with the magnification where the cell size matches with the size
displayed in the cell scale bar. *1
If the cell is defocused due to changing the objective, perform 4
again to bring the cell into focus.
For procedures to confirm the cell size while looking at the cell scale
bar, see page 53.
5
a
Focus indicator
Cell scale bar
*1Set the size of one cell to the size
between "MIN" and "MAX" from the left
end of the cell scale bar.

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If the submenu is not opened, click [Configuration] bto open the
submenu.
Click cof [Area of Culture Vessel] to display the items, and select
the area of the culture vessel to be used.
Items to be displayed are described below.
If [Custom] is selected, input the numerical value in the dfield.
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No. Type of culture vessel [mm2] (Area)
1 35mm Dish 962
2 60mm Dish 2,827
3 100mm Dish 7,854
4 150mm Dish 17,671
5 T-25 Flask (Initial setting) 2,500
6 T-75 Flask 7,500
7T-160 Flask 16,000
8 6-well 950
9 12-well 380
10 24-well 190
11 48-well 95
12 96-well 32
13 Custom Direct input
Click ein [Objective Magnification] to display the items, and
select the magnification of the objective.
Items to be displayed are described below.
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No. Magnification
1 X2
2 X4 (Initial setting)
3 X10
Click the [Count] check box fto tick it.
Click [Calibration] g. The [Cell Count Setting] screen appears.
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b c d
g
f e

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CKX-CCSW
While viewing the cell image displayed in the preview area, adjust the
threshold on the [Cell Count Setting] screen so that the point (shown
in red as default) is displayed one by one in the cell.
There are two ways to adjust the threshold: one is to click hor drag
i to adjust the slider and the other is to input the numerical value
into jdirectly.
When the threshold is determined, click [OK] k. The [Cell Count
Setting] screen is closed.
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On the [Main] screen, set the number of images to be measured while
acquiring. In [Number of Images], input the numerical value into the
lfield directly or click [+] or [-] in the mfield.
The more the number of images is increased, the higher the
measurement accuracy becomes, but it takes longer time. It
is recommended to start from approx. 5 images as a guide
and set the number of images according to the measurement
conditions.
Click [Start] n. The [Measurement Result View] screen appears.
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TIP
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h h
j
ki
ml n

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Bring the cell into focus again according to the procedure in 4.
Click [Snap] oon the [Measurement Result View] screen. The image
is acquired and the measurement results are displayed in p.
Move the culture vessel with your hand to change the acquisition
position, and repeat 14 and 15 .
Each time you acquire the image, the value displayed in pchanges.
· Single: Displays the cell count of the image acquired last.
· Average: Displays the average of the cell count of images
acquired so far.
· Estimation: Displays the estimation of the cell count of the
entire culture vessel calculated from the average.
When the number of image acquisitions set in [Number of Images] is
finished, the [FINISH] button appears.
The value in pwhen the [FINISH] button is pressed is the final result.
Click [FINISH].
The measurement results are saved and the [Measurement Result
View] screen is closed.
This is the end of the measurement of cell count.
To select the procedures to save measurement results (CSV
file or overlay image), see page 44.
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TIP
p
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