Lonza CytoSMART 2 User guide

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CytoSMART™ 2 System Manual
Bioscience Solutions

Content
1 The CytoSMART™ 2 Technology 4
2 Getting Started 5
2.1 What’s in the Box 5
3 Installing the CytoSMART™ 2 Device 6
4 Setting up Your Tablet 8
4.1 Setting up the Internet Connection 8
4.2 Starting a Project 10
4.3 Ending a Project 11
4.4 Running an Offline Project 11
4.5 Cell Counting 12
5 CytoSMART™ Connect Cloud 13
5.1 Opening Project Email 13
5.2 CytoSMART™ Connect Project Page 13
5.3 Image Analysis Settings 16
6 Troubleshooting 18
6.1 No Internet Connection Available 18
6.2 Cannot Connect with CytoSMART™ 2 Device 18
6.3 Cannot Charge Tablet 19
6.4 Video is Shaking 19
6.5 No Project Link Received 19
6.6 Cleaning the CytoSMART™ 2 Device 19
6.7 Removing Dust Particles from Cover Glass 19
7 Technical Specifications 20
8 Warnings 21

WiththeCytoSMART™2System,aneweraofpersonal andaffordablelive
cell imaging systems begins. With the integrated cloud functionality,
cell culture monitoring can occur outside of the lab, in your office or at
home, outside a clean-room or an incubator with defined (e.g. hypoxic)
conditions. Simply view your cell culture online in real-time through
the cloud with any browser-capable device, including your computer,
smartphone, or tablet.
How it Works
Setting up your CytoSMART™ 2 System does not require intensive
training, and can be set up in minutes. The CytoSMART™ 2 System
contains a mini-microscope roughly the size of a cell culture plate
fitting into any incubator. This device is linked to an accompanying
tablet that is fixed outside the incubator. Via this tablet, you can start
CytoSMART™ Projects and capture images of your cell culture. The data
is then transmitted to the CytoSMART™ Connect Cloud via wireless
transmission (WiFi) or Ethernet (LAN).
For each CytoSMART™ Project started, you will receive an email link that
allows you to monitor your cell culture remotely and in real-time.
All image files are intermediately stored on the tablet, in case
your internet access is interrupted, to assure full data safety. The
CytoSMART™ Connect Cloud is hosted via a German cloud hosting
service that currently fulfills some of the world’s strictest and most
secure data storage criteria.
Images and time-lapse videos of your cell culture can easily be
downloaded via the CytoSMART™ Connect Cloud as single images (jpg
files) or as movie files (avi files).
Applications for the CytoSMART™ 2 System
With the CytoSMART™ 2 System your cells can remain in their defined
conditions inside the incubator while you monitor the culture online.
You can also set automatic email alerts so you only need to physically
tend to your cells once they have reached a specified confluency.
Examples of Applications
– Migration assays such as scratch assays
– Documentation of stem cell culture such as reprogramming into
iPSC or for mesenchymal stem cells
– Monitoring and documentation of cell culture under restricted
conditions such as in clean-rooms or under hypoxic cell culturing
conditions
– Determination and documentation of cell growth curves
– Studies to determine influences of various factors on cell culture
performance (e.g. compound addition, screening of culture media
or growth factors)
1 The CytoSMART™ 2 Technology
www.lonza.com/cytosmart
Support Teams
Europe
Scientific Support: [phone]
[e-mail]
North America
Scientific Support: + 1 800 521 0390 (toll free)
[e-mail]
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2.1 What’s in the Box
Ensure the following items are included:
– Manual
– Tablet
– CytoSMART™ 2 Device
– Power adaptor
– Power cord
– Cable clips (3 pieces)
– License key
– USB Ethernet hub
License Key
123456789
Tablet hybrid
USB Ethernet hubLicense key
Power adaptor and
power cord
CytoSMART™2 Device
(incl. attached cable)
2 Getting Started
5
Cable clips (3 pieces)

Items required for installation:
– Incubator
– CytoSMART™ 2 Device
– Tablet
– Power adaptor
– Power cord
– Cable clips
Cable Placement
Check the incubator for a cable opening. If your incubator has one,
thread the CytoSMART™ Cable through the hole. See Figure 1. If no
opening is present, place the CytoSMART™ Cable between the door
and the frame of the incubator. See Figure 2.
3 Installing the CytoSMART™ 2 Device
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Figure 1: Placement in incubator with cable opening Figure 2: Placement in incubator without cable opening

Attachment of Tablet to Incubator Door
The tablet can be attached to the incubator with the Velcro tape in the tablet
box. To do so, peel off the plastic strip of the loop side and stick the tape to
the back of the tablet. Peel off the plastic strip of the hook side. To remove the
tablet from the keyboard base, hold the keyboard and lift one corner of the
tablet to release it. Hold the tablet with the USB port facing the door hinges
(see Figure 3). Press the tablet firmly against the incubator door.
Alternatively, the tablet with attached keyboard can be placed beside the
incubator. Tablet can be closed during a running project as power safe mode
of tablet is disabled.
USB Connection
The cable connected to the CytoSMART™ 2 Device should be connected to the
tablet. Connect the power adaptor to the tablet and the power cord. Plug the
power cord into the electrical outlet (see Figure 4).
Secure Cables
The cables can be secured to the incubator using the three cable clips
provided. It is advised to use one cable clip to secure the connection of the
CytoSMART™ Cable to the tablet. This can be done by placing the cable clip just
behind the USB connector that goes into the tablet (see Figure 5). This will
ensure the connector stays in position when the incubator door is opened.
Indicator Light
To confirm the system is running properly turn on the tablet, open the
incubator and check to see if the light panel around the CytoSMART™ 2 Device
is illumitated:
– Blue light: no project running
– Red light: project running
T-ask Placement
The CytoSMART™ 2 Device accomodates a variety of different culture vessels,
including T-flasks, well plates, single dishes and flat tubes.
It is advised to place the CytoSMART™ 2 Device in the incubator with the LED
holder pointing to the left. This way the live image on the tablet moves in the
directions indicated by the arrows when culture vessels are moved on the
CytoSMART™ 2 Device as shown (see Figure 6).
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Ethernet / LAN
Ethernet / LAN Connection
If WiFi is not present you can connect the CytoSMART™ 2 Device to the internet
using an Ethernet hub (see Figure 7). It is important to follow a specific
connection sequence: Switch on the tablet, disconnect the power adaptor from
the tablet if it is connected, connect the power adaptor with the Ethernet hub,
then connect the Ethernet hub to the tablet. If notification “Please connect
charger” appears, repeat the above steps. Connect the Ethernet cable to your
LAN.
Important: To connect the CytoSMART™ 2 Device to the Ethernet, only use
the Ethernet hub provided with the CytoSMART™ 2 System.
Figure 3: Attachment of tablet to incubator
Figure 5: Secure cables
Figure 4: Connection
Figure 6: T-flask placement and positioning
Figure 7: Ethernet hub connection

Figure 8: Power adaptor and power cord for charging tablet
4 Setting up Your Tablet
IMPORTANT: You must have the CytoSMART™ Cable connected
to the tablet and the tablet connected to a power outlet.
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To set up your system, have the following available:
– Login details of a stable WiFi internet or LAN connection (no
guest network)
– CytoSMART™ 2 Device
– Tablet (connected to power source)
– A valid license key
4.1 Connecting to the Internet
Charging the Battery
Fully charge the tablet battery before using it.
Connect the power adaptor to the tablet and to the power cord. Then
connect the power cord to a pow er outlet.
The power cord varies by country or region. Make sure to only use the
power adaptor and power cord that is supplied in the package. Using a
different charging unit or cable may damage your tablet.
Turning on the Tablet
To turn on your tablet, hold down the power button on the top edge for
a few seconds, then release it.
The tablet manual can be downloaded from the manufacturer’s
website.
Connecting to the Internet
After the CytoSMART™ 2 Software loads, a yellow bar with the message
“Please connect to the internet.” will appear at the top of the screen. To
proceed you first need to connect the tablet to the internet via a WiFi
or LAN connection.
To connect the tablet to the internet swipe from right to left on the
outside of the right side of the screen. Select “Network” (1).
Select your WiFi network (2) and log in.
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How to Minimize the CytoSMART™ App
Use the minimize button to get to the Windows desktop, enabling
access to Windows features and applications like Edge (internet
browser). To minimize the CytoSMART™ App, select the “Menu” button
(3) followed by the “Minimize” (4). To go back to the CytoSMART™ App,
select the application in the task bar.
Starting the CytoSMART™ App
The CytoSMART™ App will launch automatically when the tablet is
turned on.
Entering the License Key
Enter the license key (5) that can be found in the CytoSMART™
Accessories box. Use either the keyboard base or the on-screen
keyboard, which can be opened by pushing the keyboard button at the
display bottom. Read terms and conditions and accept by checking
the check box.
You are now ready to start a project (6).
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4.2 Starting a Project
Place a culture vessel containing cells on top of the CytoSMART™ 2
Device. You should now see the cells on the screen.
Setting Focus
Use the slider (7) to adjust the focus. The plus and minus buttons
can be used for more precise adjustments.
Zoom Functionality
To digitally enlarge the image, use the “Zoom” function. Select “Menu”
then select “Zoom In”. To return the image to standard formatting,
select “Zoom Out”.
Start Recording
To start a recording, select “Project” (8).
Entering a Project Name
Select the “Project Name” field (9) and give your project a name.
Please note that the Project Name field is mandatory. Use either
the keyboard base or the on-screen keyboard which is opened by
pushing the keyboard button at the display bottom. You can add an
Extra Note (10) if desired (e.g. cell type, passage, etc.). Select “Next”.
Entering Your Email Address
Next, enter your email address to receive the project link (11). Select
“Next” (12).
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IMPORTANT: Please ensure the email address is correct, other-
wise you will not receive the email with a link to the project.
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Selecting the Recording Frequency
Select the desired recording frequency (13). Use the drop down menu
to select an interval between 1 and 60 minutes. Please remember that
biological cells can be susceptible to light. It is advised not to use a
higher recording frequency than needed.
Starting the Project
After you select the “Start” button (14) you will receive an email with
a link to open your project. Your system will immediately enter sleep
mode. Tap anywhere on the screen to re-activate the screen.
When the CytoSMART™ 2 Device is operated by the tablet with attached
keyboard, the tablet can be closed during a running project. The power
safe mode of the tablet is disabled in plugged-in and battery mode to
prevent the tablet from shutting down.
4.3 Ending a Project
To finish a project select the red “Stop project” button (15). The system
will send you an email indicating that the project has ended.
4.4 Running an Oine Project
The CytoSMART™ 2 System can be operated in offline mode. Set the
tablet to flight mode to run a project in offline mode. If keyboard
is attached, select the key with an airplane symbol on it, otherwise
swipe from right to left on the outside of the right side of the screen.
Select “Network” (16). On the next screen select “Flight mode” (17).
To start a recording, select ”Start project” [18]. Give your project a
name, select a snapshot interval, then select “Start”.
Snapshots from offline projects are stored under:
C:\ProgramData\CytoSmartApp\Images\”. To browse to this folder,
please select “Menu” followed by “Browse”. The CytoSMART™ App will
minimize automatically and a window with the Images folder will open.
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Please note: The CytoSMART™ 2 Device measures and displays
the temperature of the device surface, which can be slightly
above the incubator average. For temperature sensitive cells,
please avoid extended monitoring in live mode or snapshot
intervals of 1 min, which can lead to an increase of up to 1.0°C.
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4.5 Cell Counting
Cell counting can be done with the CytoSMART™ 2 System by selecting
„Count“ in the „Menu“ (19). Cells in suspension (adherent cells after
having been detached from their culture vessel or cells growing in
suspension) are to be diluted (typically 1:2) with Trypan Blue (use
a ready-to-use 0.4% solution, available from many suppliers). Ensure
the cells are in single cell suspension.
Pipette the suspension into a counting slide or chamber. Be certain the
height of the chamber or the distance between slide and cover glass is
0.1 mm (typical of most counting slides or chambers).
Place the slide or chamber on the CytoSMART™ 2 Device so cells are
in an area without grid lines. It is important to adjust the focus so
the cells are visible as small white dots with black rims (20). Use the
Zoom-in function for more precise focusing.
Select “Menu” (19) followed by “Count” (21). The counting and
calculation take approximately 20 seconds. Only the cells in the center
part of the field of view will be counted and shown in the subsequently
appearing display. The counted live cells are noted in green, the
counted dead cells noted in red.
CytoSMART™ 2 System counting results are presented as number
of live cells per milliliter, number of dead cells per milliliter, total cell
number per milliliter, and viability (% live cells of total cells) (24).
Record the CytoSMART™ 2 System results and ensure you multiply
them by the Trypan Blue dilution factor to determine final results.
Select „Return” (23) to revert to the live image again.
Selecting „Browse“ (22) in the „Menu“ will reveal an Explorer window
(25) containing the „Counting“ folder (26). Two snapshots for each
cell counting are stored here; one showing the cell results noted in
green and red (27), and one without the color markings (28).
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To connect to the CytoSMART™ Connect Cloud you will need:
– A device able to run an internet browser (e.g. computer, smart
phone or tablet)
– An internet connection
– CytoSMART™ Project Email
Once you start a project, the CytoSMART™ 2 System will begin to
capture images of your cell culture. Images are then transmitted to
the CytoSMART™ Connect Cloud.
5.1 Opening Project Email
For each CytoSMART™ Project started you will receive an email
containing a summary of your project and a link to the CytoSMART™
Connect Cloud (See Figure 9).
To access your project, select the “View” button in the email. This will
take you to your CytoSMART™ Connect Project Page where you can
monitor your cell culture remotely with any browser-capable device.
5.2 CytoSMART™ Connect Project Page
Overview
An overview of all the panes and options on the CytoSMART™ Connect
Project Page and detailed information on individual features.
From: Lonza Bioresearch Solutions [mailto:[email protected]om]
Sent: Thursday, 6 July, 2017 14:23
To: Technical Support - Verviers <[e-mail]>
Subject: New Project Started: 2017-07-06_AoSMC_CytoSMART 2
A new project started at 12:23 on Thursday 6th July 2017 UTC(London Winter
Time)
. Please keep this email as it contains the access key for your cell culture's
information. You can view, edit, or export details by clicking on the "View" button.
2017
-07-06_AoSMC_CytoSMART 2
Project Name:
2017-07-06_AoSMC_CytoSMART 2
Project Note:
Project Owner:
scientific.support[email protected]
Project Start Date:
06-07-2017 12:23:27 UTC(London Winter Time)
Duration:
-
Average Temperature:
-
Snapshot Interval:
15 minutes
Snapshot Count:
-
You have received this email from
http://www.lonza.com/research because our system is
monitoring a cell culture. This email contains information about
the cell cultivation and you may be required to take action.
Figure 9: CytoSMART™ Project Email
5 CytoSMART™ Connect Cloud
IMPORTANT: If you do not receive an email, please ensure you
entered the correct email address and check your spam folder.
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https://cytomate.com/access/Project_main/main.php?DiD=117&PiD=1942&ac1=4624e7df56e027625dfe32c78c8f0501634d1387fe863af72971a7b4a1b53fdb&a…
1/2
[e-mail]
01d:15h:23m | 05d:22h:22m
Graph Edit
Cell Coverage
Details
Time: 01d:15h:23m Cell Coverage: 76.0%
Temperature: 37.0°C Snapshot No.: 312 (11 Jan 2015 07:38)
Temperature
Summary
Project Start Date:
Project Note:
Project Owner:
Duration:
Fri, 9 Jan 2015
1+DF
MRKQ[e-mail]
5d:22h:22m
Average Temperature: 36.98°C
Snapshot Interval: 7.5 minutes
Snapshot Count: 1128
Notifications Edit
To set notification, press the edit button...
Cell Coverage
Coverage over: 70%
Temperature
Temperature over: 39°C
Logbook
Notification Added 21-01-2015 15:10
Notification Added 21-01-2015 15:10
Project Ended 15-01-2015 14:40
Project Started 09-01-2015 16:15
Notes
Enter a Note...
Save
Video View Tracking
Follow us on: Privacy PolicyTerms & Conditions© 2015 CytoMate Technologies BV
Project: B15W0003
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38.2°C
37.9°C
37.7°C
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0 1d 4h 2d 8h 3d 13h 4d 17h 5d 22h
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1. Download movies (.avi), images (.jpeg), cell coverage, and temperature values (.csv).
2. Option to view the cells analyzed by the image analysis algorithm in the time-lapse video.
3. Time-lapse cell culture video.
4. Video control buttons (play, pause, forward one picture, backward one picture).
5. Slider to control movie (point in time).
6. Editing button to change image analysis settings.
7. Indicator line showing point in graph corresponding to video time point.
8. Cell coverage graph showing result of image analysis.
9. Details box showing exact values of Time, Temperature, Cell Coverage and Snapshot number associated with specific time points.
10. Temperature graph
11. Control to speed up, or slow down the time-lapse video.
12. Summary pane
13. Notification pane
14. Logbook pane
15. Notes pane
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Summary Pane
The summary pane provides an overview of your project (i.e. project
start date, duration, average temperature and number of images
taken (snapshot count)).
Notication Pane
In the notification pane you can set thresholds to enable the system
to notify you if:
– Cell coverage reaches a specified percentage
– Temperature reaches a specified limit
To set a notification, select “Edit”. The notification pane will unfold
allowing you to specify the type and value of notifications. You can
add multiple upper and lower limits and notifications can be deleted
or added at any time during your project. Notifications will be sent to
the email address entered when the project was started.
Logbook Pane
The logbook pane provides a in front of historic log of all changes
made to your CytoSMART™ 2 System.
Notes Pane
You can add additional information (e.g. passage number, experiment
conditions, etc.) in the notes pane. The information will be date and
time stamped and stored in the project database
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5.3 Image Analysis Settings
Depending on the size and morphology of your cells it may make
sense to change the image analysis settings. You can do so by
changing the tracking settings to a different grade. This will alter the
algorithm used to calculate confluency.
To edit the image analysis settings, select the “Edit” button (16)
in the graph under the time-lapse video. A dialog box that enables
changing the tracking settings will appear.
Tracking Settings
Select a new tracking grade in the drop down menu (17). After
selecting a grade, the system will provide a preview of your selected
snapshot. By selecting the checkbox “Tracking” (18) you can switch
between the tracking images and the original images.
Grade Selection
Choose the tracking grade that best fits your cells. You can select
a predefined tracking grade (20) by selecting the drop down menu
(17).
If none of these options fit your desired tracking grade, please select
“-Custom-“ in the drop down menu or select the “More” button (19).
You can use the sliders to create custom settings. The green markings
of recognized cells will adapt directly to any slider change.
The tracking grade consists of three parameters:
Block size (Number of Pixel) (21):
– The smallest element of the image that is individually
processed
– The smaller the block size is, the more detailed cell structures
are recognized as part of a cell
Threshold (22):
– Adjustment of cell recognition versus background
– Lower thresholds result in greater cell recognition
Cap level (23):
– Lower cap levels result in more structures being identified as
cells
With the slider (24) you can navigate to different project snapshots.
The tracking grade will be refreshed automatically. Cell coverage is
shown for the current snapshot (25).
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If tracking grade settings are changed, the previous cell coverage will
be shown in brackets (26).
Select “Apply” (27). This will re-run the image analysis. If “Cancel”
(28) is selected changes will not be saved.
Select “OK”. It will take a few moments for the settings to be updated.
Processing
The system is now re-calculating the image analysis. The system
re-calculates approximately 1000 snapshots per minute.
Refreshing your Web-browser
Your web-browser can store the old CytoSMART™ Connect Project
Page in the cache memory. To ensure you see the results of the re-
calculated image analysis you should force your web-browser to
refresh. Depending on your operating system all you need to do is
press the following key combinations:
– Windows: F5
– Mac OS X: <cmd> + R or Apple + R
– Linux: F5
Make sure to push the buttons simultaneously.
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IMPORTANT: The time required for processing depends on the
duration of the recordings. Long time-lapses will require addi-
tional time.
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6.1 No Internet Connection Available
If you receive a warning indicating you do not have an internet
connection (1), please try the following:
– Check to see if your tablet is connected to a network via WiFi or
Ethernet (see Chapter 4).
– Reboot your tablet. Minimize the CytoSMART™ App by “Menu”
followed by “Minimize”. Touch the “Windows” symbol (2) and
select the switch off button (3). Select “Restart” (4).
– Verify the WiFi or LAN connection has access to the internet by
opening the CytoSMART™ Website www.lonza.com/cytosmart.
This can be done by minimizing the CytoSMART™Application
(see above) and selecting the browser on the task bar. Please
ensure the website is loading. If the page is not loading, verify
the WiFi or LAN connection on another device (PC, smart phone
or tablet) with a working internet connection. If it is working
correctly, contact your IT department for further assistance in
accessing a network connection for your device.
6.2 CannotConnectwithCytoSMART™ 2Device
If you receive a warning indicating the system can not connect with
the CytoSMART™ 2 Device (5), this is usually the result of the USB
cable not being well connected, thus preventing the CytoSMART™ 2
Device from communicating with the tablet. To resolve this please
try the following:
– Make sure the CytoSMART™ 2 Device is connected to the tablet.
– Switch off the tablet. Unplug and re-connect all cables. Reboot
the tablet.
6 Troubleshooting
Please connect to the internet
1
Please connect the device
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6.3 Cannot Charge Tablet
A warning indicating the tablet is not charging (6), is typically the
result of either the power adaptor not being well connected, the power
cord not having good connection in the power outlet, or the power
outlet being turned off. To resolve this please try the following:
– Make sure the power outlet is functioning (e.g. check by plugging
in another electrical device).
– Use a different power outlet.
– If USB Ethernet hub is connected, unplug it from the tablet. Unplug
the power adaptor from the USB Ethernet hub. Plug the power
adaptor to the USB Ethernet hub and then plug the USB Ethernet
hub to the tablet.
6.4 Video is Shaking
If images in your time-lapse recording appear to be shaking it is likely
due to a problem with the bearings of the incubator fan or a nearby
shaker (e.g. in a bottom incubator). To check this please place the
CytoSMART™ 2 Device outside of the incubator and start a project to
see if the shaking persists.
6.5 No Project Link Received
The CytoSMART™ 2 System requires a robust internet network
connection (WiFi or LAN) to function properly. Faulty or slow internet
networks can cause delays in email notifications, image submission
to the cloud and project link transmission. If you’ve waited several
minutes and still do not receive a project link after you have started a
project, please do the following:
– Check to see if you entered the correct email address
– Check to see if your SPAM filter blocked the email
Alternatively, you could select an alternate email address with
different SPAM filters (e.g. Gmail, Hotmail).
6.6 Cleaning the CytoSMART™ Device
The CytoSMART™ 2 System requires minimal maintenance for reliable
operation. Disconnect the CytoSMART™ 2 Device from the tablet before
cleaning and disinfecting the case. Use a damp cloth to wipe down the
outer case with water or 70-80% ethanol.
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IMPORTANT: Do not autoclave the CytoSMART™ 2 Device.
Autoclaving will result in damage to the electronic system.
Please connect charger
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6.7 Removing Dust Particles from Cover Glass
If dark spots are visible in the displayed image in the absence of
microscopy objects they may be caused by dust and other particles
on the cover glass. Remove them by gently wiping the cover glass with
a soft cloth.

7 Technical Specications
Feature Technical Data
Power Supply AC 100-240V, 3A, 15W, 50/60Hz
Field of View 2.4 x 1.4 mm
Magnication 10X
Image Resolution 1280 x 720 pixels (converted to 640 x 360 on webpage)
Exported Formats JPEG, CSV, AVI
Light Source LED
Camera 5 MP CMOS
Display Remote 10” tablet touchscreen
Optical Filters No optical lters
Unit Dimensions 133 x 90 x 100 mm (LxWxH)
Weight 0.5 kg
Data Storage 50 GB online
Operating Environment 5–40 °C, 20–95% humidity
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