CURIOSIS Celloger Nano User manual

www.curiosis.com
Automated live cell imaging system
Celloger Nano™
Instruction manual

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Developed and manufactured by CURIOSIS Inc.
Email: [e-mail]
Website: www.curiosis.com
CURIOSIS Inc.
4F, 10 Teheran-ro 38-gil, Gangnam-gu, Seoul 06221, South Korea
Tel: [phone]
Fax: [phone]
Copyright © 2021, by CURIOSIS Inc.
All rights reserved. Published in Korea.

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Table of contents
Package contents 4
Device layout 5
Getting started 6
Pre-requirements 6
Installation 7
Software description 10
1)Celloger Nano APP 10
2)Celloger Nano Analysis APP 17
Operation overview 18
Create or open a project 19
Device connection 20
General operation 21
1. Preparation (cell/device preparation) 21
2. Cell monitoring and image setting 22
2.1. Location adjustment 22
2.2. Light intensity adjustment 26
3. Image analysis 27
3.1. Capture 27
3.2. Intensity 29
3.3. Ruler and manual count 30
4. Time-lapse 31
5. Process single image or time-lapse images 33
5.1 Import the original data taken by celloger nano 33
5.2 Slide show 34
5.3 Merge fl image 34
5.4 Estimate confluency (Coverage) 35
5.5 Make video 36
Safety instruction 37
General guidelines 37
Operating condition 37
Safety standards 38
Cleaning and maintenance 38
Trouble shooting 38
Product specifications 40
Ordering information 41
Appendix A. Connecting multiple devices 42
1. Installation 42
1.1 Device ip setup 42
1.2 Using a router 43
2. Operation 44
Appendix B. Alternative PC network setup 45

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Package contents
Celloger NanoTM package includes the following items.
Item
Quantity
Celloger Nano
1
Main Power Cable
1
POE Injector
1
Adapter
1
LAN Cable (5M, 4.8M)
2
USB memory
1
- Instruction Manual
1
- Celloger Nano App
1
- Celloger Nano Analysis App
1
When receiving the package,
• Check that all items listed above are included in your package.
• Examine the device carefully for any damage during shipping.
• Contact your local distributor or inf[email protected] if any items are missing or damaged.
• Any loss or damage claims must be filed with the carrier.

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Device layout
Front-left side
POE Injector

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Getting started
Pre-requirements
Installation location
For normal and stable operation of the device, the following environmental conditions should be satisfied.
Keep the device at temperature between 10 ~ 40 °C.
It is not recommended to operate the device at low temperature (Below 10 °C).
In those conditions, warm up the device for over 10 minutes.
Relative humidity between 0 ~ 95 %.
Install it on a flat and level surface.
Place at dust- or other airborne particles-free area.
During operation, place around 10cm away from the surroundings for proper air flow of the cooling fan.
During operation inside incubator, do not place close to the incubator door.
Do not put any heavy materials on top of the device.
Do not place it together with vibrating equipment.
Upon using multiple equipment, there should be a proper clearance among equipment.
PC specifications (Refer to page 40)
Celloger Nano software can be used on PCs with Window 10.
Large capacity hard drive is necessary to save images from Timelapse. If the capacity is not enough,
scanning will not function properly.
For the ease of use, it is recommended to use a monitor with resolution of 1920 x 1080.
LAN port is required to connect the equipment to PC using LAN cable. If there is no LAN port, LAN cable
adapter should be prepared.

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Installation
1. Prepare a computer that meets the specifications and place it near the incubator. (PC specifications
p.40)
2. Take the equipment out of package and put it on a flat desk.
3. Connect two LAN cables and power cable to the POE injector.
Connect one LAN cable to the equipment (yellow) and another LAN cable to the PC (white).
4. Connect the power cable to an outlet.
5. Turn on the power by pressing the power switch located next to the power cable port.
6. Perform the PC network setting (changing PC IP address) as follows to connect the PC with the
equipment. Alternative method to set PC network within the Celloger Nano App can be found in
Appendix B (p.45)
6.1 Click the “Start” button or the “Window” icon.
6.2 Click the “Setting” button.
6.3 Click “Network and Internet” in Control Panel.
6.4 Click the “Ethernet” button.
6.5 Click “Network and Sharing Center”.
6.6 Click “Change adapter settings” on the left-side menu.
6.7 Select “Ethernet” (This name can vary according to PC.) button and right-click the mouse and click
“Properties” in the window that appears.
6.8 Select “Internet Protocol Version 4 (TCP/IPv4)” and click “Properties”.
6.9 Select “Use the following IP address” and enter IP address (192.168.2.XX*) and Subnet mask
(255.255.255.0) in the blank fields
Note. Fill 2~254 except 10 in XX* field.
- The PC IP address should be set differently from the
device IP address or network adapter such as a
router. The default device IP is 192.168. 2. 10 and to
set up the PC IP address, enter 192.168.2.XX (in XX
field, input 2~254 except 10).
- This IP setting guideline may not be applicable to
some PC depending on the PC network environment.
In case of connection failure, contact us via
[e-mail] or local distributor.
6.10 Click OK, then network configuration is
completed.
Power cable
LAN cable into device
LAN cable to PC

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7. To start Celloger Nano App, first, connect the USB drive provided with the device into the PC, then
copy APP folder to the PC and execute “Celloger Nano App.exe” in the APP folder.
<Image Descriptions of PC network setting>
①
②
③
④
⑤
⑥

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⑦
⑧
⑨

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Software description
Celloger Nano provides two software: Celloger Nano App and Analysis App. Celloger Nano App is a program
for actual operation of Celloger Nano device that includes real-time monitoring and time-lapse imaging
function. Celloger Nano Analysis App is a program for analyzing or post-processing the images obtained from
Celloger Nano and it operates even without the device connection.
1) Celloger Nano APP
Upon opening Celloger Nano App and completing pre-setup (Refer to p.19~20), the following main page is
displayed.
①Menu Bar
[ File ]
✓Open Project : Import a previously saved project.
✓Save Project : Save the settings on the current project.
✓Save Project As… : Create a new project with the current settings (Light
intensity, Focus position, Schedule, etc.).
✓Save Image As… : Save the Preview image.
①
②
③
④
⑥
⑤

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[ Tool ]
[ Device ]
[ Help ]
②Toolbox
[ Preview ]
: Streaming on/off on the Display. The live image is shown based on the selected channel such
as bright- field (Hereafter ”BF”), green fluorescence (Hereafter “FL green”) and red fluorescence (Hereafter
“FL red”) at the “Light Source Control”.
✓ : Streaming on
➢Visibility : Show/hide features displaying image information such as
histogram.
✓Histogram (p.29) : Show/hide intensity histogram of an image.
✓Intensity (p.29) : Show/hide point and mean intensity of an image.
✓Manual Counting : Show/hide “Manual Counting” function.
✓Scale Bar : Show/hide a scale bar.
✓Center Mark : Show/hide a cross-shaped feature at the center of the
display for reference.
✓Connect : Connect a new device or change the connection to another device.
✓Reboot : Reboot the connected device.
✓IPAddress : Change the IP address of the connected device.
✓Status : Display the status of connected device.
✓Parameter : Display the values set for the connected device. Certain
parameters can only be manipulated by an authorized person.
➢Manual Counting (p.31) : Count the points clicked by user.
✓Count : Add a point for counting.
✓Remove : Delete a clicked point.
✓Clear : Delete all points.
➢Reset Layout (p.13) : Return to User Interface before change
➢Preferences : Set “AutoFocus” (Refer to p.24), merge layer (Refer to
p.28), and appearance (change the color and font size of the feature).
✓About CellogerApp: Display App information.
✓ : Streaming off

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[ File ]
: Save the image on the Display.
[ Zoom & Pan ]
✓ : Zoom in (Digital zoom)
✓ : Zoom out (Digital zoom)
✓ : Pan (Drag the display in Zoom-in status)
✓ : Return the zoomed image or relocated image to its original state at once.
[ Measure ]
Measure the length between the two clicked points.
✓ : Measure the distance between two selected points.
✓ : Measure the distance between two selected points on horizontally.
✓ : Measure the distance between two selected points on vertically.
[ Layer ]
✓ : Merged layer (Merge activated layers of BF and FL image).
Note. Once activating “merged layer”, the displayed image is an image processed by setting
“Opacity” and “Threshold” in the “Preferences” (Refer to p. 28).
✓ : BF image layer
✓ : FL (FL Green/FL Red) image layer
✓ : Features [Ruler(Measure), scale bar, manual count, etc.] layer
[ Device control ]
✓ : Execute autofocusing in streaming on state according to autofocusing setting (Refer to p.24).
✓ : Turn on the white light in the LED status indicator of the connected device.
✓ : Fan on/off
③Display
Show real-time image upon activating Preview or the last captured image during time-lapse.

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④Light Source Control
The tabs in “Light Source Control” are composed of BF, FL green and FL red. The configuration of tabs are
set depending on the light options of Celloger Nano.
✓LampPower (%) : Control lamp (Illuminator) power (0~100%).
✓Exposure (Second): The duration of exposure of cells to light (0.010~2.000 second). Be careful as
photobleaching occurs when exposure value is high.
✓Gain : The amount of signal amplification (1.0~10.0) during the process of changing photon data to
digital data. This is a parameter associated to the contrast of image. Be careful as noise becomes
higher when gain value is high.
✓Offset : (Black level) Brightness of a dark range of image. Be careful as contrast becomes low when
offset value is high.
⑤Stage & Time-lapse Control
Window can be separately floated so that it can fit on the monitor. It is possible to change the window by
pressing (Context Menu) for floating or hiding. It can convert to AutoHide/Dockable immediately by
clicking . The Layout is automatically saved upon terminating the program. To go back to original state,
press “Reset Layout” in the “Tool” before closing the program.
Note. When the window disappears, delete “LayoutCNN.xml” in the program folder to go back to default
state after closing the program.
[ Z Stage control ]
A. Current Z stage coordinate
B. Upon pressing “Go” after entering the desired coordinate in the box, the stage is moved according
to the set coordinates. Upon pressing , current coordinate is entered in the Box.
C. Distance moved upon pressing Jog button (D) once (Hereinafter referred to as “step distance”).
: Step distance unit : Step distance
A
C
B
D

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D. Jog button
: Move Z stage up. [Z coordinate + step distance]
: Stop the stage on the move.
: Move Z stage down. [Z coordinate - step distance]
[ Focus ]
Select the channel used for time lapse imaging and select the focus position (Z coordinate) for each channel.
✓Imaging Channel : Channel used for time-lapse imaging. BF is always checked.
✓Ref. Pos. (Reference position)/ Offset
➢Reference position : Position of BF channel for time-lapse.
A. : Upon clicking, the current coordinate is entered on the box next to it.
B. : The location of z axis of BF channel when Auto Focus function is off (None) or median
coordinate when Auto Focus is on (FirstCycle or EveryCycle).
C. : Upon clicking, location moves according to coordinates shown on the left box.
➢Offset from BF reference position : Position of FL channel for time-lapse.
A. : Upon clicking, the current coordinate is entered on the box next to it.
B. : The location of z axis of FL channel captured while performing time lapse. The value
shown is the difference from the location of Z axis captured in the BF channel(B). To capture the
FL channel on the same coordinates of BF channel, enter 0.
C. : Upon clicking this button, move to coordinates shown on the left box.
A
B
C
D
E
F

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✓Auto Focus
[ Schedule ]
Set the schedule for time-lapse imaging.
Time except for End Date and Total Cycle can be selected. Click using the mouse, turn the mouse wheel or
click right arrow or calendar for selection.
✓Interval time : Interval between scanning cycles
✓Total Running Time : Time-lapse execution period
✓Start Date : Time lapse start time
(If the time set is past time, the time changes automatically to the time when the button is pressed upon
pressing “Start”.)
✓End date : Time lapse complete time
(It is determined by Total Running Time and Start Date, and the user cannot directly change the value.)
✓Total Cycle Expected :The number of scans during time lapse period
(It is determined by Interval time and Total Running Time, and the user cannot directly change the value.)
Note. The total cycle expected is calculated by simply dividing the total running time by interval time and
does not consider the data processing time. Therefore, the expected total cycle could be different from
the actual scanning cycles.
➢None : During time-lapse imaging, it does not execute autofocusing function and
captures based on the entered coordinates (Ref. Pos./Offset B)
➢FirstCycle : During time-lapse imaging, the autofocusing function is executed only
for the first cycle and the capturing for other cycles is done on the z coordinates
designated in the first cycle.
➢EveryCycle : During time-lapse imaging, execute autofocusing function every
cycle.

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[ Time lapse panel ]
A. The time elapsed (“Elapsed”) and progress rate (%) during the total time lapse running time. Count
means the number of times scanning occurred. Upon clicking Start, scanning begins and this refers
to count 1. (Count = Cycle count + 1)
B. The progress rate (%) within a cycle and remaining time until the next cycle (“Remaining”). Index
shows the number of images taken within a cycle (With Celloger Nano, Index is always 0).
C. Current date and time.
D. Upon clicking “Start”, scanning starts based on the schedule.
⑥Connection status
It shows device connection status, and information on connected devices.
A. IP address of connected device.
B. Description of connected device (Can be changed in “Device”-> “Parameter”).
C. Serial number of connected device.
D. Firmware version of connected device.
Note. If device is not connected, shows up. Click “Device”-> “connection”
for reconnection. If the connection window does not show the device icon, Refer to p.20.
A B
C
D
A
B
C
D

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2) Celloger Nano Analysis APP
Upon opening Celloger Nano Analysis App and selecting ‘Config file’, the following main page is displayed
①Menu bar
It is managed by “config file” instead of a project.
Other menus are the same as those of Celloger Nano App.
②Toolbox
: Mask the area recognized as confluency (Possible to use after confluency analysis). All other tools
and functions are the same as those of Celloger Nano App.
③Display
Display the image selected from Image Analysis control list (⑤). The image is displayed based on the
activated channel that can be selected in “Light source control” (④). Upon completing analysis, coverage
mask is shown and functions such as measure (Ruler), Intensity (point), manual counting can be
performed on the screen.
①
②
③
④
①
②
③
④
⑤

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④Light Source Control
Display the light source settings of Celloger Nano App and it cannot be changed.
⑤Image Analysis Control
Upon importing a folder, file list is displayed.
Upon selecting the file, the image is shown on
Display.
It is possible to analyze images or make video.
(Refer to p.33)
Analyze: Calculate the confluence of the files
checked on the list. Analyze the image of
selected light channels on the right (Refer to
p.35).
Record: Make video (Refer to p.36).
Play: Show the images in the list in order (Refer
to slideshow p.34).
Stop: Stop playing.
Operation overview

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Create or open a project
✓With Celloger Nano, the settings can be managed by the project file (“.cgnproj” file format). Setting values
(Light intensity, Focus position, Schedule, file storage location, etc.) are saved in the project.
✓Upon executing App, popup screen for project selection is displayed.
✓Create a new project or open and existing one. You can start a program with default project by clicking
“Continue without configuration”.
✓Upon selecting Create Project, popup window that requires the selection of project name and storage
location shows up. You can start the program after entering required information and press OK. The
storage location of the project is the address of default program file and cancel “Use default location” and
enter address to change the storage location.
Import the project saved in the
past.
Create new Project.
Start the program without
Customizing a Project. In this case,
a default project “Experiment” is
executed.

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Device connection
Icon of devices connected by LAN cable is shown. To connect the device, select the icon and press “Connect”.
Upon completing the connection, the color of LED status indicator changes from red to green.
Note. If the icon is not shown, check the following steps and take appropriate actions.
Case 1. PC IP setting is not completed properly.
Reset the PC IP. If the PC IP, Device IP, Router IP are the same, connection fails (PC IP setting p.7 step 6).
Case 2. LAN or power cable is not connected properly (LED status indicator is turned off).
Check all cable connection status and POE injector power (Installation p.7 step 3 to 5).
Case 3. Separate LAN Adaptor is used.
Press the arrow on the right to select the
adaptor to which the device is connected.
Case 4. When using multiple devices in one PC, each device has the same IP.
Initially, devices are released with the same IP. When using the device for the first time, it is necessary to reset the IP address.
(Appendix. A. Device IP setting p.42)