Azure Biosystems Azure Cielo Dx AIQ314 User manual

Azure Cielo™ Dx
Real-Time PCR Systems
User Manual
Part Numbers AIQ314 and AIQ315
UM-0008 R3

Azure Cielo Dx Real-Time PCR Systems User Manual Page 2
Safety and Regulatory Compliance
Important Safety Information
Please read these instructions before operating the Azure Cielo Dx system.
Electrical Safety Precautions
Be sure to take proper precautions when handling any electrical equipment. NEVER work on any live
circuit, xture, receptacle, or switch. Safety rules you should follow whenever working with any electrical
appliance include:
• Always shut off power at the main disconnect before changing a fuse.
• Always shut off power to the circuit before repairing or replacing a switch, receptacle, or xture.
• Always tape over the main switch, empty fuse socket, or circuit breaker you are working on.
• Always check that the circuit is dead before beginning work on it. Using a circuit tester or voltmeter
can help you determine this.
• Always unplug any appliance before repairing it.
Protective ground terminal
The ground terminal, intended for connection to an external protective conductor for protection
against electric shock in case of a fault, is located on the inside of the back panel.
NOTE: This equipment has been tested and found to comply with the limits for a Class A digital device,
pursuant to Part 15 of the FCC Rules. These limits are designed to provide reasonable protection against
harmful interference when the equipment is operated in a commercial environment. This equipment
generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the
instruction manual, may cause harmful interference to radio communications.
Operation of this equipment in a residential area is likely to cause harmful interference in which case the user
will be required to correct the interference at his own expense.
Changes or modications not expressly approved by the party responsible for compliance could void the
user’s authority to operate the equipment.
Hot surface warning
Under normal conditions, the temperature of plate is below 50°C and safe to touch. However, if
the system malfunctions, it is possible that the glass surface temperature exceeds 80°C. Please
exercise caution when touch the plate surface with hand when this occurs.
CAUTION: No user serviceable parts inside!

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For in vitro diagnostic (IVD) use
The Azure Cielo 3 Dx and Azure Cielo 6 Dx Real-time PCR systems are registered
with the U.S. FDA as Class II 510(k) exempt devices.
Europe: For IVD use. The Azure Cielo 3 Dx and Azure Cielo 6 Dx systems are CE
marked under the In Vitro Diagnostic Medical Devices Directive (98/79/EC). The CE
IVD–authorized Azure Cielo Dx Systems are for distribution and use the following
countries: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czechia, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia,
Liechtenstein, Lithuania, Luxembourg, Malta, the Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, and the
United Kingdom.
Incident Reporting
Any serious incident that has occurred in relation to the device shall be reported to the manufacturer and the
competent authority of the Member State in which the user and/or the patient is established.
Warranty
The Azure Cielo Dx products are warranted against defects in materials and workmanship for one year
unless otherwise outlined on your sales order or www.azurebiosystems.com/warranty. If any defect occurs in
the instrument during this warranty period, Azure Biosystems, Inc. will repair or replace the defective parts at
its discretion without charge. The following defects, however, are specically excluded:
• Defects caused by improper operation.
• Repair or modication done by anyone other than Azure Biosystems or the company’s
authorized agent.
• Use of spare parts supplied by anyone other than Azure Biosystems.
• Damage caused by accident or misuse.
• Damage caused by disaster.
• Corrosion caused by improper solvents or samples.
Voltage Setting Information
The Azure Cielo Dx system has a power supply that automatically chooses the correct voltage for your
country or region.
CE Conformity
The following Cielo Dx Model Systems, models: Cielo 3 Dx and Cielo 6 Dx are in conformity with
the provisions of the following EC Directives, including all amendments, and national legislation
implementing these directives:
• EN61010-1: 2010
• EN61326-1: 2013
• EN61326-1:2013
• EN301489-1 v2.2.1:2019
• EN301489-17 v3.2.0:2017
• EN300238 v2.1.1:2016
Protection category: IP20 according IEC 60529

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ENG
This symbol indicates that the waste of electrical and electronic equipment must not
be disposed as unsorted municipal waste and must be collected separately. Please
contact an authorized representative of the manufacturer for information concerning
the decommissioning of your equipment.
FRA
Ce symbole indique que les déchets relatifs à l’équipement électrique et électronique
ne doivent pas être jetés comme les ordures ménagères non-triées et doivent être
collectés séparément . Contactez un représentant agréé du fabricant pour obtenir des
informations sur la mise au rebut de votre équipement.
GER
Dieses Symbol kennzeichnet elektrische und elektronische Geräte, die nicht mit dem
gewöhnlichen, unsortierten Hausmüll entsorgt werden dürfen, sondern separat behandelt
werden müssen. Bitte nehmen Sie Kontakt mit einem autorisierten Beauftragten des
Herstellers auf, um Informationen hinsichtlich der Entsorgung Ihres Gerätes zu erhalten.
ITA
Questo simbolo indica che i riuti derivanti da apparecchiature elettriche ed elettroniche
non devono essere smaltiti come riuti municipali indifferenziati e devono invece essere
raccolti separatamente. Per informazioni relative alle modalità di smantellamento delle
apparecchiature fuori uso, contattare un rappresentante autorizzato del fabbricante.
SPA
Este símbolo indica que el equipo eléctrico y electrónico no debe tirarse con los
desechos domésticos y debe tratarse por separado. Contacte con el representante local
del fabricante para obtener más información sobre la forma de desechar el equipo.
SWE
Denna symbol anger att elektriska och elektroniska utrustningar inte får avyttras
som osorterat hushållsavfall och måste samlas in separat . Var god kontakta
en auktoriserad tillverkarrepresentant för information angående avyttring av
utrustningen.
Waste Electrical and Electronic Equipment (WEEE)
Contact
Azure Biosystems, Inc.
6747 Sierra Court, Suite A-B • Dublin, CA 94568 • USA
[e-mail] • [phone] • Fax [phone]

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Table of Contents
1. Introduction 6
1.1 Azure Cielo Dx Specications 6
1.2 Plates and Tube Compatibility 7
1.3 Contacting Technical Support 7
2. Installation and Setup 8
2.1 System Placement 8
2.2 System Overview 8
2.3 Connect Azure Cielo Dx Real-Time PCR system to Internet Network 9
2.2 Connect Azure Cielo Dx Real-Time PCR system to PC via
Azure Cielo Dx Manager software 9
3. Device Software Overview 10
3.1 The Home Screen 10
3.2 Experiment Setup 10
3.2.1 Protocol Manager 11
3.2.2 Change Protocol Properties 13
3.2.3 Well Selection Screen 15
3.2.4 Email Feature in Protocol Setting 15
3.2.5 Run Setup 16
3.2.6 Run Protocol 21
3.3 Data Manager 23
3.4 Device Settings 27
3.4.1 General 27
3.4.2 Network 27
3.4.3 Email Server Setup 28
3.4.4 Dye Manager 30
3.4.5 Technical Support 30
4. FAQs and Troubleshooting 31
4.1 FAQs 31
4.2 Troubleshooting 31

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1. Introduction
The Azure Cielo Dx Real-Time PCR instrument is a high performance tool compatible with a wide variety of
reagents for qPCR. The family of instruments includes the Azure Cielo 3 Dx and the Azure Cielo 6 Dx.
1.1—Azure Cielo Dx Specications
The Azure Cielo Dx instruments have the following specications.
Specications Azure Cielo 3 Dx
Real Time PCR System
Azure Cielo 6 Dx
Real Time PCR System
Part number AIQ314 AIQ315
Product description
96-well Real-Time PCR instrument
with 10.2” touchscreen interface,
3 dye channel lters
96-well Real-Time PCR instrument
with 10.2” touchscreen interface,
6 dye channel lters
Sample capacity (wells) 96 96
Reaction volume 10–150µL 10–150µL
Excitation source LED LED
Detection channels 3 6
Multiplexing Up to 3 targets Up to 6 targets
Thermal element Peltier Peltier
Max. block ramp rate 6°C/sec 6°C/sec
Avg. sample ramp rate 4°C/sec 4°C/sec
Temperature uniformity ±0.2°C ±0.2°C
Temperature accuracy ±0.1°C ±0.1°C
Dye compatibility
SYBR Green, EvaGreen, FAM,
VIC, JOE, HEX, CAL Fluor 540,
CAL Fluor Orange 560, Cy5, LIZ,
Mustang Purple
SYBR Green, EvaGreen, FAM,
VIC, JOE, HEX, CAL Fluor 540,
CAL Fluor Orange 560, ROX,
TAMRA, TEX615, Quasar 670,
CAL Fluor Red 610, Cy5, LIZ, Mustang
Purple, Cy5.5, Quasar 705
Custom dye/chemistry üüüü
Chemistry capability Fast/Standard Fast/Standard
Detection sensitivity 1 copy 1 copy
Sensitivity Detect differences as small as 2-fold in
target quantities in singleplex reactions
Detect differences as small as 2-fold in
target quantities in singleplex reactions
Connectivity USB, Wi-Fi, Ethernet USB, Wi-Fi, Ethernet

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1.2—Plates and Tube Compatibility
Azure Cielo Dx Real-Time PCR system is compatible with:
A. Low-prole Plates
• 96-well, Either Non-skirted or Semi-Skirted
• Low-prole 96-well Plates: These types of plates have wells of approximate height 15.5 mm
(depending on manufacturer). These are modern designs that have the potential to reduce
condensate formation and offers the advantage for capturing light in uorescence assays,
low-volume reactions, and fast PCR.
• Non-skirted Plate: This plate lacks surrounding panel. This format ts blocks of most thermal
cyclers and qPCR systems, but it is not suitable for robotic applications.
• Semi-skirted Plate: This plate has a short panel around the edge of the plate, providing
adequate support during pipetting and mechanical strength for robotic handling.
• Color of the well can be White, Frosted or Clear
• Plates should be sealed with Optical Grade Seal Film
• Plate rim (lip): The “rim” of a qPCR plate is a peripheral upper elevation around the plate. The
qPCR plates with rim are NOT compatible with the Cielo Dx system.
• Plate Notch: The notch is a modied corner of a qPCR plate, which is determined by the
instrument being used for optimal t. The notch maybe at the four corners of the 96-well
plate at positions A1, H1, A12 and H12. Cielo Dx instruments are compatible with any
notch positions.
B. Low-prole Tubes
• Either individual or 8-strip or 12-strip
• Tube formats: Tubes are an ideal choice for qPCR experiments of low to medium throughputs.
Individual and tube strips are two most common formats available. Tube strips are available
in 8 or 12 well type. Cielo Dx systems are compatible with 0.1ml and 0.2ml.
• Color of the well can be White, Frosted or Clear
• The tubes should be sealed with Optical Grade Flat Caps
• Tube caps: Cap strips are the most common form of qPCR caps to seal tube strips and 96-
well plates. In addition to t, clarity and shape are two main features to consider in choosing
the caps. Azure Cielo Dx Real-Time PCR systems are compatible with Flat Caps, that are
Optical or Ultra-Clear.
1.3—Contacting Technical Support
For questions about installation, setup or general use of the system, please contact
[e-mail] or call [phone], 9am–4pm Pacic time.

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2.1—System Placement
WARNING: Excessive Weight Hazard – Please use two or more people to lift the system. Failure to do so
can result in system damage and personal injury.
As with all electrical instruments, the Azure Cielo Dx should be located away from water, solvents, or
corrosive materials, on a at and stable surface with adequate clearance on all sides. The system must
remain stationary during operation.
The system should be placed away from interfering electrical signals and magnetic elds. If possible, a
dedicated electrical outlet should be used to eliminate electrical interference from other instrumentation in
your laboratory.
The Azure Cielo Dx should be installed at no more than 3000 meters above sea level.
2.2—System Overview
2. Installation and Setup
Figure 2. Side view of the Azure Cielo Dx
Real-Time PCR systems.
Figure 1. Front view of the Azure Cielo Dx Real-Time PCR systems.
Touch screen panel
USB Type A port
Door
Start button with indicator LED
Open button
Figure 3. Back view of the Azure Cielo Dx
Real-Time PCR systems.
Power Switch
Power Input USB Type A port
Ethernet port
USB Type B port

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2.3—Connect Azure Cielo Dx Real-Time PCR system to Internet Network
A. Wi-Fi
1. Go to the DEVICE SETTINGS.
2. Select Wi-Fi, under the NETWORK settings on the left edge of the screen.
3. Make sure that the WiFi is turned on.
4. Find your network from the populated list.
5. Input your password and encryption type.
6. Then the system should connect to the WiFi network.
B. Ethernet
1. Connect a LAN or ethernet cable to the Ethernet port at the rear of Azure Cielo Dx Real-Time
PCR system.
2. Go to DEVICE SETTINGS > NETWORK > Ethernet to make sure connection is properly established.
Note: Depending on your institution’s network security settings, you may need to provide the Mac Address
shown on your Cielo 6 Dx to your IT department to add it into the whitelist of your internet network. You
can nd the Mac Address from DEVICE SETTINGS > NETWORK > Wi-Fi > CONNECTION INFO.
2.4—Connect Azure Cielo Dx Real-Time PCR system to PC via Azure Cielo Dx Manager software
A. Internet connection
1. Please make sure both Azure Cielo Dx Real-Time PCR system and the PC are connected to the
same internet network (please refer to Section 2.3).
2. Install Azure Cielo Dx Manager software on the PC with administrative privileges.
3. On the startup wizard, click “SEARCH DEVICES” button to search available Azure Cielo Dx
Real-Time PCR system, then click “CONNECT” to connect selected system to the PC.
B. USB connection
1. Connect Azure Cielo Dx Real-Time PCR system to a PC by using USB Type B cable (a USB Type B
to USB Type A cable).
2. Plug the Type-B connector into the USB Type-B port at the rear of Azure Cielo Dx Real-Time PCR
system, and the Type-A connector to the standard USB port on the PC.
3. Install Azure Cielo Dx Manager software on the PC with administrative privileges.
4. On the startup wizard, click “SEARCH DEVICES” button to search available Azure Cielo Dx
Real-Time PCR system, then click “CONNECT” to connect selected system to the PC.

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The Azure Cielo Dx Icon based software allows you program and run protocols with a simple user interface.
3.1—The Home Screen
3. Device Software Overview
Figure 4. The Home Screen.
Button Button Name Function
Experiment Setup Set up a new run or running an existing protocol –
see Section 3.2
Data Manager Access run data – see Section 3.3
Device Settings Access device settings – see Section 3.4
Connection Bar Status of internet connection
Table 1. Buttons in the home screen.
3.2—Experiment Setup
In short, to run a qPCR protocol, rst create new user or protocol, or select existing user and
protocol in PROTOCOL MANAGER screen. Then, tap go to PROPERTIES screen to edit
protocol properties.
Next, tap go to RUN SETUP to edit run setup. Finally, tap to run the protocol, the
information of real-time run will be displayed on the RUN screen.

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3.2.1—Protocol Manager
Figure 5. PROTOCOL MANAGER screen
under Experiment Setup.
Button Button Name Function
Home Back to Home Screen
Connection Bar Status of internet connection
Sort Sort in ascending or descending order
Add • Add User
• Add Protocol
Delete • Delete User
• Delete Protocol
Next Go to the PROPERTIES screen to edit the protocol properties
after selecting an existing or newly created protocol
Table 2. Icon buttons in the PROTOCOL MANAGER screen.
Figure 6. PREVIEW screen of a selected
protocol in PROTOCOL MANAGER.

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A. Tap button next to Users or Protocol and Date created to sort users and/or experiments in
ascending or descending order.
B. Add New User:
• Tap button to add a new user.
• Tap on ‘Add User’ to add a new user. Enter the new user name using the system
keyboard.
• You can see the new user added to the PROTOCOL MANAGER screen.
Figure 7. Add New User
process ow.
C. Add New Protocol:
• Tap symbol to add a new protocol.
• Tap on ‘Add Protocol’ to add a new user. Enter the name for the protoocol using the
system keyboard.
• You can see the new protocol added to the PROTOCOL MANAGER screen. The PREVIEW
screen of a new protocol displays the default experiment settings.
Figure 8. Add New Protocol
process ow.
Figure 9. The PREVIEW screen of a new protocol
displays the default experiment settings

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D. Delete exisiting user or protocol.
• Tap on the User or Protocol to be deleted.
• Tap icon to delete the selected User or Protocol.
Note: Deleting user will delete all existing protocol and data stored under that user.
• The system will prompt you to conrm the deletion. Tap ‘OK’ to delete.
Figure 10. Delete exisiting user or
protocol in PROTOCOL MANAGER.
3.2.2—Change Protocol Properties
1. Tap to select the protocol to edit properties.
2. Tap icon to edit the selected protocol.
Figure 11. Select a protocol in PROTOCOL MANAGER.
Figure 12. PROPERTIES screen of a selected protocol.

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Button Button Name Function
Home Back to Home Screen
Connection Bar Status of internet connection
Protocol Name Tap the rectangular text box to assign a name (or rename)
the selected protocol
Reaction Volume
• Select qPCR reaction volume
• Select OTHER button to customize
reaction volume
Lid Temperature Select lid temperature
Off lid heating Turn off lid heating
Light Source Select light source(s)
Back Back to PROTOCOL MANAGER screen
Next Go to RUN SETUP screen
Email Send run completed notication with result
(.AZD le) attached
Well Selection Go to WELL SELECTION screen to select wells from
a 96 well PCR plate
*Selected button will turn green from default blue color.
Table 3. Icons and buttons in PROPERTIES screen of a selected protocol.

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3.2.3—Well Selection Screen
• A screen with a diagram of 96 well PCR plate appear on the screen. By default, all wells are
selected (in BLUE).
• To deselect wells, tap on the well to clear selection. Wells are deselected in sets of two
columns. Press to save changes on the well plate and return to Properties screen.
Example: Deselected Column 1, 2 and 11, 12.
Note: 2 column of wells are deselected per tap.
• You may also use ‘CLEAR ALL’ and tap to select the wells. Press to save changes on the
well plate and return to PROPERTIES screen.
Note: 2 column of wells are deselected per tap.
• Tap to proceed to RUN SETUP screen.
Figure 13. WELL SELECTION screen. Figure 14. Deselected columns 1,2 and 11,12 in
WELL SELECTION screen.
Figure 15. Select only Column 11 and 12 after “CLEAR ALL” selection.
3.2.4—Email Feature in Protocol Setting
Important: Make sure that:
1. Cielo Dx device is successfully connected to the Internet. To connect the Cielo Dx Device
to the internet, please refer Section 2.3 (Connect Azure Cielo Dx Real-Time PCR system to
internet network).
2. Email server is successfully setup. To set up email server, please refer Section 3.4.3
(Email Server Setup)

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Figure 16. Email ID pop-up screen.
• A pop-up screen with an empty column appear allows user to input email address.
• Input email ID and click OK.
• Proceed with subsequent Experiment Setup and run protocol.
• Email notication with results attached will be sent when the protocol run is
successfully completed.
3.2.5—Run Setup
Figure 17. RUN SETUP screen.
Button Button Name Function
Home Back to Home Screen
Connection Bar Status of internet connection
Melt Curve On/Off
• Turn on Melt Curve by tapping the switch button, then
select the temperature resolution
• When BLUE, the Melt Curve is ON; When GRAY, the Melt
curve is OFF
Melt Curve
Advance Setting
Melt Curve Advance Settings: starting temperature, ending
temperature, resolution, and scan function

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Button Button Name Function
Temperature Tap to customize reaction temperature
Time Tap to customize time in the step
Cycle Setting Tap to assign total cycle number and the step number to
repeat every time after the last step of the cycle
Add a step Tap on this icon to select the step after which a new step
must be inserted
Delete a step Tap on this icon to select the step which must be deleted
Go To module Duplicate existing cycling steps in the RUN SETUP
Save Saving Run Setup Settings
Camera and/or
Gradient PCR Advance settings on camera and/or gradient PCR
Back Back to PROPERTIES screen
Run Run the protocol
1. Temperature settings:
• Tap on the temperature display button for each cycle to change the temperature(°C). Enter
the numerical digits [Example: 60] using the key pad on the screen. Press ‘DONE’ to save.
• Tap to clear the entered numerical digits.
Table 4. Icons and buttons in RUN SETUP screen of a selected protocol.

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2. Time settings:
• Tap on the time display button for each cycle to change the time. Enter the time using the
key pad on the screen.
Example: To set the time to 2 minutes, Tap ‘Min’ on the screen and enter the number 2
using the key pad on the screen. Press ‘DONE’ to save.
Example 2: To set time to 1 minutes 30 seconds, tap on ‘Min’ on the screen and enter the
number 1. Tap on ‘Sec’ and enter the number 30 using the key pad on the screen. Press
‘Done’ to save.
• Tap to clear the entered numerical digits.
3. Cycle Settings:
• Tap on the purple display button under the CYCLING stage. The number on the
button displays the number of cycles.
• Tap on to enter the number of cycles you want to run.
• Tap on to enter the step number you want the cycle to repeat everytime, after the
last step of the cycle.
Example: To set 40 cycles consisting of step 2 and 3, tap on the purple button that
displays number of cycles. Tap on CYCLES and enter 40. Tap on GO TO, and enter 2
(Step number).
4. Go To module:
• Tap the Go To cycling step then the button will duplicate the existing cycling steps
in the RUN SETUP. User can freely add or delete steps with the and button, and
assign temperature and time for these new steps.
Figure 18. Go To module on the
RUN SETUP screen.
5. Add or delete a step:
• Add a step: Tap to select the step after which a new step must be inserted.
• Delete a step: Tap to select the step which must be deleted.

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6. Advance settings on camera and/or gradient PCR:
A. Camera:
• By default, the camera is disabled (LIGHT GRAY ). Tap to enable the camera for the
desired steps, tap on settings . Tap on the switch button next to CAMERA to turn
the camera ON. Tap to save changes and to return to the RUN SETUP
screen without saving changes.
• The switch button turns BLUE when the camera is ON and GRAY when the camera
is OFF.
Figure 19. Advance settings screen
for camera and/or gradient PCR.
• The camera at a particular step is active if the icon display is DARK GRAY.
Example: In the image left, the camera at STEP 3 and melt curve are active.
B. Gradient PCR:
• To enable the gradient of the PCR step, tap on settings . Tap the switch button next
to GRADIENT to turn the gradient ON (the switch button will turn BLUE when gradient
is ON). Tap to save changes and to return to the RUN SETUP screen
without saving changes.

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Example: In the following image, gradient is ON for Step 2 and 3 and OFF for Step 1
and 4. Gradient can be turned on for all steps, individually.
7. Melt Curve Advance Settings:
• Tap the switch to turn on Melt Curve .
• Tap to access advance setting for Melt Curve.
• Adjust starting temperature, ending temperature, resolution, and scan.
• Tap to save the modied Melt Curve advance settings.
Figure 20. Melt Curve advance settings screen.
8. Saving Run Setup Settings:
• Tap to save the current settings to the protocol. This options enable the user to directly
load and repeat the same experiment again at a later time.
9. Run Experiment: Real Time
• Tap to run the saved protocol.