vivoVerse vivoCube+ User manual

Document Number: VVS-QA-016-02
[e-mail]
vivoCube+ User Manual™
v1.1, Hardware Rev B – August 2023

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© 2023 vivoVerse, Inc. All rights reserved.
/
Disclaimer
This instrucon manual is for the vivoCube+ system. To ensure safety, obtain opmum performance,
and to become suciently familiar with the use of this device, any user of the vivoCube+ should study
this manual thoroughly before operang the vivoCube+. Retain this instrucon manual in an easily
accessible place near the vivoCube+ for future reference. If the equipment is used in a manner not
specied by this manual, the safety of the user and he funconal integrity of the vivoCube+ may be
compromised. Always use the vivoCube+ as outlined in this instrucon manual.
Intended Use
This device has been designed to be used as a pressure source in various roune lab work and research
applicaons. Do not use this device for any purpose other than its intended use. This device is
intended solely for research use and is not to be used for any other purposes including, but not limited
to, unauthorized commercial purposes (including Contract Research, diagnoscs, or manufacture),
military purposes, veterinary or clinical use. Contact vivoVerse for full terms and condions.
Handling Precauons, Maintenance, and Storage
•The vivoCube+ is a precision scienc instrument. Use reasonable care in using, handling,
storing, transporng, and disposing of the vivoCube+ (including by wearing appropriate
protecve equipment). Ensure it is used only by qualied laboratory personnel who have been
trained to use it. Avoid subjecng it to sudden or severe impact.
•Connect any cables or tubing to the vivoCube+ gently.
•Do not use the vivoCube+ where it is subjected to direct sunlight, high temperature, high
humidity, dust, or strong vibraons.
•Do not disassemble any part of the vivoCube+.
•To clean ngerprints, dirt, or stains, use a piece of cleaning paper or ssue moistened with
commercially-available, aqueous, mild cleaning soluon.
oDo not use acetone or alcohol to clean the vivoCube+.
•When disposing of this product, follow any regulaons and rules on the disposal of electronics
set by your organizaon and/or the local government.
Warranty
Visit vivoverse.com/salesterms for limited warranty coverage for the vivoCube+.
Copyright
All rights reserved. This manual or any poron thereof may not be reproduced or used in any manner
whatsoever without the express wrien permission of the author except for the use in brief quotaons
in a review. © 2023 vivoVerse, Inc.
vivoVerse
3303 Northland Dr Suite 308
Ausn, TX, 78731
[e-mail]

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User Manual Changelog
Version
Publish Date
Version Notes
v1.0
2022-06-14
First revision of the vivoCube+ manual for Hardware Rev A,
vivoCubeOS v1.0
v1.0.1
2022-08-18
Fixed typos. Added note about inspecng tubing for wear and
cauon that it is connected properly.
v1.1
2023-10-01
Updated to vivoVerse branding. Updated features and
instrucons for vivoCubeOS v1.1.5 and Hardware Rev B.

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© 2023 vivoVerse, Inc. All rights reserved.
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Table of Contents
1. Introducon ........................................................................................................................................... 5
1.1. Specicaons .............................................................................................................................. 5
2. vivoCube+ Basics.................................................................................................................................. 8
2.1. Front Panel................................................................................................................................... 8
2.2. Rear Panel .................................................................................................................................... 9
2.3. Pneumac Connecons..........................................................................................................10
2.3.1. Using preconnected tubing with luer lock connectors ..............................................10
2.3.2. Using push-to-connect ngs on the vivoCube+ rear panel ..................................11
2.4. vivoCube+ Power.........................................................................................................................12
2.5. vivoCube+ Quality Inspecon...................................................................................................12
3. User Interface ......................................................................................................................................13
3.1. Home....................................................................................................................................13
3.2. Manual.................................................................................................................................14
3.3. Programs ..............................................................................................................................15
3.3.1. Channel Selecon Menu..................................................................................................16
3.3.2. Program Progress Menu ..................................................................................................17
3.4. Sengs .................................................................................................................................19
3.5. Info ........................................................................................................................................21
4. Upcoming Features.............................................................................................................................22
5. Potenal Issues and Troubleshoong .............................................................................................23
Appendix......................................................................................................................................................25
A. Using an External Pressure Source...........................................................................................25
B. Updang vivoCube+ Soware..................................................................................................25
C. vivoCubeOS changelog...............................................................................................................30
D. Reporng Bugs and Subming Feature Requests................................................................31

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1.Introducon
vivoCube+ is a congurable pressure-driven uid controller
designed to drive steady, smooth ow of liquids. The vivoCube+ can
be used with custom-programmed pressure sequences or a library
of preset programs ready for immediate use.
The operang owrates and pressures are specied so that
vivoCube+ can be used not only for C. elegans experiments1 using
vivoVerse vivoChip-2x devices but also for a wide variety of other
microuidic applicaons:
•Organoids and 3D organotypic cultures
•2D organ-on-a-chip models
•Zebrash models
•Drosophila larvae
•Other worm models
•Single cell models
1.1. Specicaons
Table 1 outlines pressure control performance specicaons for the
vivoCube+.
Table 1: vivoCube+ specicaons for pressure control performance.
Performance Specs
Pressure range (built-in pump)*
0 to 10 psi (gauge)
Pressure range (oponal
external pressure source)
0 to 30 psi (gauge)
Sensing accuracy
±0.1 psi
Sensing resoluon
±0.002 psi
Control accuracy
±7% from 0.1 to 0.5 psi
±2% from 0.5 to 1 psi
±1% greater than 1 psi
Control stability
±0.01 psi
Maximum ow rate (built-in
pump)*†
1000 mL/min
1 Every vivoCube+ is pre-
programmed with
patented pressure cycles
developed to immobilize
C. elegans within a
vivoChip-2x for high-
resoluon imaging
* Congurable upon
request
† total ow rate is divided
among the two channels
if operated
simultaneously

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The specs outlined in Table 1 are dened as follows:
•Pressure range (built-in pump): the output pressure range
that the pump built into the vivoCube+ can provide at zero
ow rate.
•Pressure range (external pressure source): the range of
pressures that can be connected to the Pext port on the
vivoCube+ and controlled via either pressure channel. Before
using an external pressure source with the vivoCube+, it is
crical to read Appendix A.
•Sensing accuracy: the range within which the average
pressure readings from the vivoCube+ pressure sensors will
fall with respect to a (hypothecal) known pressure value.
•Sensing resoluon: the minimum incremental pressure
change that the pressure sensors built into the vivoCube+
can detect.
•Control accuracy: the range within which the average
pressure readings from the vivoCube+ pressure sensors will
fall with respect to a desired pressure setpoint set by the
user. Reported value is based on a representave load
connected to the vivoCube+; control accuracy is ulmately
applicaon-dependent and will vary.
•Control stability: the range within which any given pressure
reading from the vivoCube+ pressure sensors will fall with
respect to a desired pressure setpoint set by the user.
Reported value is based on a representave load connected
to the vivoCube+; control stability is ulmately applicaon-
dependent and will vary.
•Maximum ow rate (built-in pump): the maximum ow rate
the built-in pump can provide.2
2 vivoCube+ is intended to
be connected to a uid
reservoir to drive uid
ow using air pressure.

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Table 2 outlines general specicaons for the vivoCube+.
Table 2: General vivoCube+ specicaons.
General Specs
Supply voltage
12 VDC,
Supply current (minimum)
1 A
Dimensions
250 mm x 155 mm x 54 mm
Weight
820 g
Independent pressure channels
2 (A and B)
Flow media compability
Non-corrosive, non-ionic gases,
such as air
Input pneumac connecon
(oponal)
1/8” inner diameter (ID), 1/4”
outer diameter (OD) tubing
(Pext)
Output pneumac connecons
1/16” ID, 1/8” OD tubing (PA,
PB)
The vivoCube+ requires electrical power to funcon. A power
adapter is provided with interchangeable N-, E-, B-, and A-type
blades shown in Figure 1, covering most power outlets in North
America, Europe, United Kingdom, and Australia, respecvely.3
Figure 1: Diagram of the dierent power blade adapters packaged with
the vivoCube+ system.
3 If you require a dierent
type of power adapter for
your electrical power
supply, please contact
[e-mail]
with your country
informaon.

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2.vivoCube+ Basics
This secon consists of a descripon of the various physical
features, controls, and indicators on the vivoCube+.
Each vivoCube+ system includes the following components:
•vivoCube+
•12 VDC, 1 A power supply with interchangeable power
blades
•USB Type-B cable (white)
2.1. Front Panel
A top-down view of the vivoCube+ is depicted in Figure 2, showing
the vivoCube+ front panel. The front panel contains the user
interface/control panel of the vivoCube+.
Figure 2: Annotated top-down view of vivoCube+ (front panel).
1. Screen: high-contrast, monochromac OLED screen that
displays menus and informaon during vivoCube+ operaon.
2. Mulfuncon dial control: primary input control for the user
to operate the vivoCube+. The dial can be rotated either
clockwise or counterclockwise to move the cursor
throughout the menus displayed on the screen. The dial can

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also be pressed to select the opon highlighted in white in
the displayed menu.
3. Ring light indicator: LED-illuminated ring that acts as an
addional visual indicator (for example, the ring briey
ashes white whenever the user selects a menu opon).
In general, navigaon through the vivoCube+ graphical user
interface (GUI) is achieved by using the mul-funcon dial control.
Rotate the dial clockwise to advance through a menu or an opon;
rotate the dial counterclockwise to step back through a menu or an
opon. Press the dial to “click” and select the currently highlighted
opon. When certain events occur, the ring light indicator might
ash, change color, or fade in or out as an addional visual
indicator.
In addion to the controls and indicators depicted in Figure 2, the
vivoCube+ contains a speaker, which produces chime sounds under
certain condions. For example, a chime plays when the pump is
powered on or o or when a program commences or completes.
Screen and ring indicator brightness sengs and sound sengs can
be modied in the Sengs menu, described in secon 3.4.
2.2. Rear Panel
A rear view of the vivoCube+ is depicted in Figure 3, showing the
vivoCube+ rear panel. The rear panel contains power, pressure, and
data ports.
Figure 3: Annotated rear view of vivoCube+.
1. Power switch: the rocker switch that powers on or o the
vivoCube+ when power is supplied through the power jack

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(Figure 3 item 6). When the top part of the switch is
depressed, a blue LED light within the switch is illuminated,
indicang that the vivoCube+ is powered on.
2. Channel A pressure port (PA): the pressure port controlled
within vivoCube+ soware as pressure channel A. 1/8” OD
tubing can be connected to this port.
3. Channel B pressure port (PB): the pressure port controlled
within vivoCube+ soware as pressure channel B. 1/8” OD
tubing can be connected to this port.
4. USB port: a USB connecon to a desktop computer is
possible with this port using the USB Type-B cable provided
with the vivoCube+. This manner of connecon is required
for vivoCube+ soware updates. More informaon on
soware updates can be found in Appendix B.
5. External pressure source port (Pext): the pressure port that
can oponally be connected to an external pressure source.
1/4” OD tubing can be connected to this port. For more
informaon on how to use an external pressure source with
the vivoCube+, see Appendix A.
6. Power jack: the socket to which the vivoCube+ power supply
(provided with the vivoCube+) should be connected. Ensure
that the power switch is in the o posion before connecng
the power supply to the power jack. Do not use a power
supply other than the one provided with the vivoCube+.
2.3. Pneumac Connecons
This secon outlines methods to connect pneumac tubing to the
vivoCube+. The vivoCube+ outputs controllable air pressure that
can be used to pressurize a liquid reservoir (such as a ported GL45
bole) to drive liquid ow.
2.3.1.Using preconnected tubing with luer lock connectors
The vivoCube+ system includes tubing connected to the Channel A
and Channel B pressure ports at the end of which is a plug/male
luer lock. For convenience and reliability of the uidic connecon, it
is recommended to use a socket/female luer connector to connect
your applicaon to the tubing connected to the Channel A and

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Channel B pressure ports. The connecon can be made using the
twist-lock mechanism of the luer lock ngs, shown in Figure 4.
Figure 4: Diagram of plug/male and socket/female luer lock connectors.
2.3.2.Using push-to-connect ngs on the vivoCube+ rear
panel
To connect tubing directly to Channel A pressure port, Channel B
pressure port, or the external pressure source port, the end of the
appropriately-sized tubing must be inserted by hand into the port
by rmly pushing the tubing into the green ng4. Check that the
tubing is inserted completely into the ng. As the tubing is
pushed deeper into the ng, resistance should be felt, indicang
that the internal mechanism of the ng is gripping the tubing and
sealing the connecon to it. Eventually the tubing will not be able to
be pushed in any farther, at which point it is properly and fully
inserted. Figure 5 shows a diagram of the push-to-connect
operaon for reference.
4 Note: improper
connecon of the tubing
to pressure port is a
common mistake in the
setup of the vivoCube+.

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Figure 5: Rear panel of the vivoCube+ with diagram of push-to-connect
ng operaon.
To disconnect the tubing, press and hold the green release ring and
then pull the tubing out of the ng.
2.4. vivoCube+ Power
To power on the vivoCube+, rst check that the power switch is in
the o posion. Then, connect the vivoCube+ power supply to the
power jack. Finally, ip the power switch to the on posion. To
power o the vivoCube+, ip the power switch to the o posion5.
2.5. vivoCube+ Quality Inspecon
The vivoCube+ package contains a quality inspecon document
(vivoCube+ Quality Inspecon Document, document number VVS-
QA-016-F01), which should be completed upon recepon of the
vivoCube+ system. Complete this document before using the
vivoCube+ for the rst me.
5 Warning: do not remove
the vivoCube+ power
supply from the power
jack while the power
switch is in the on
posion.

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3.User Interface
Throughout the menus of the vivoCube+ graphical user interface
(GUI), in general, a highlighted secon of the screen (whether
highlighng a menu opon, buon, or other item) indicates the
current cursor posion. The cursor posion can be changed by
turning the dial on the front panel of the vivoCube+ by hand.
Clockwise rotaons advance the cursor posion forward through
menus while counterclockwise rotaons go backward through
them. Pressing down on the dial and “clicking” it results in a user
selecon within the GUI. Dial rotaon and clicking are the means
for navigang the vivoCube+ GUI6.
When the vivoCube+ is rst powered on, a loading screen appears,
during which user sengs are loaded into the main program. Upon
compleon of the loading process, the Home menu is displayed.
3.1. Home
The Home menu is depicted in Figure 6 with the three menu
opons and other indicators annotated. The Manual opon is
highlighted, indicang that the cursor is currently posioned there.
Figure 6: Annotated drawing of Home menu.
1. Current menu indicator: in this case, Home is displayed
here. For all menus throughout the vivoCube GUI, the current
menu is displayed in the middle of the top of the screen as
shown in Figure 6.
6 The easiest way to learn
how to use the vivoCube+
GUI is to explore the
menus by following on the
vivoCube+ while reading
this secon.

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2. Manual: clicking on this opon will bring the user to the
Manual menu, which allows on-demand, manual control of
each pressure channel. More informaon on the Manual
pressure control menu can be found in secon 3.2.
3. Programs: clicking on this opon will bring the user to the
Programs menu, which lists the various pressure programs
available to run7. A number of preset pressure programs are
included with each vivoCube+. These programs involve
applying a sequence of dierent pressure values for dierent
duraons of me. More informaon on the Programs
menu and programs in general can be found in secon 3.3.
4. Sengs: clicking on this opon will bring the user to the
Sengs menu, which lists several operaonal sengs that
the user can modify. This menu also includes system
informaon, such as serial number and soware version.
More informaon on the Sengs menu and programs in
general can be found in secon 3.4.
5. Descripve text: short line of text outlining the funcon of
the currently highlighted menu opon.
3.2. Manual
The Manual menu is depicted in Figure 7 with the various menu
opons and indicators annotated.
Figure 7: Annotated drawing of Manual menu.
7 The vivoCube+ is pre-
programmed with
programs for every
vivoVerse vivoChip-2x
model. Use the
corresponding program
with the chip being
operated, and follow the
on-screen instrucons for
best performance.

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1. Set P(A): set channel A pressure. Clicking this opon brings
the cursor to the channel A pressure setpoint (Figure 7 opon
5).
2. Toggle P(A) (“Turn on P(A)”/“Turn o P(A)”): this opon will
read either “Turn on P(A)” or “Turn o P(A)” depending on the
current on/o state of the pressure output. Turning on P(A)
will bring the output pressure in channel A to the setpoint set
in Figure 7 opon 5. Turning o P(A) will block output
pressure from the vivoCube+, bringing it to 0 psi output;
however, the setpoint for P(A) will not change8.
3. Set P(B): same funconality as Set P(A) but for channel B
pressure.
4. Toggle P(B) (“Turn on P(B)”/“Turn o P(B)”): same funconality
as Toggle P(A) but for channel B pressure.
5. Channel A pressure setpoint: clicking Set P(A) (Figure 7
opon 1) will move the cursor to the channel A pressure
setpoint, allowing the user to change the pressure setpoint
digit by digit, starng with the ones place. Click through each
digit to set the pressure to the desired value; all four digits
must be set to a number between 0 and 9. Aer the last digit
is clicked, the cursor will return to Set P(A).
6. Channel B pressure setpoint: same funconality as Channel A
pressure setpoint but for channel B pressure.
7. Channel A pressure current value: a live reading of the actual
pressure measured at the Channel A pressure port (PA).
8. Channel B pressure current value: same funconality as
channel A pressure current value but for channel B pressure.
9. Home: clicking this brings the user to the Home menu9.
10. Programs: clicking this brings the user to the
Programs menu.
11. Sengs: clicking this brings the user to the
Sengs menu.
3.3. Programs
The Programs menu is depicted in Figure 8 with the menu
opons annotated. In this depicon, the cursor is posioned at the
rst menu opon. As the user advances through the menu, the
programs list scrolls, displaying more programs.
8 Tip: using the Toggle P(A)
and Toggle P(B) opons
allows the user to pulse
output pressure on
demand.
9 Warning: leaving the
Manual menu (via Figure 6
opons 9-11) turns o the
pump and resulng
pressure control—stay
within the Manual
menu if maintaining a
desired pressure is
needed.

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/
Figure 8: Annotated drawing of Programs menu.
1. Programs selecon menu: a list of default programs available
to run is displayed and able to be scrolled through on this
menu. Select a program to proceed to run it; selecng a
program brings the user to the channel selecon menu,
described in secon 3.3.1.
2. Home: clicking this brings the user to the Home
menu10.
3. Manual: clicking this brings the user to the Manual
menu.
4. Sengs: clicking this brings the user to the Sengs
menu.
oNote: if a program is currently running, the opon
Now Running will be displayed instead of Sengs.
This allows the user to cancel the selecon of a new
program and return back to the currently running
program.
3.3.1.Channel Selecon Menu
Whenever a program is selected from the Programs menu, the
user must select which channel the program should be applied to as
shown in Figure 9. In this example, the program that was selected
was “D1-D5 vivoChip-2x.” Whichever channel is selected in this
menu is the one that will be set up to run this program on the
Program Progress menu (secon 3.3.2).
10 Warning: leaving the
Programs menu (via
opons 2-4) turns o the
pump and any currently
running programs—stay on
the Programs menu or
associated program status
menu to ensure the
compleon of any
selected programs.
Secon 3.3.2 contains an
excepon to this rule for
the scenario in which a
program is complete.

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Figure 9: Channel Select menu (within Programs menu).
Note that channel selecon can be aborted in the Channel
Selecon menu by selecng Programs and that the Programs
menu can be exited enrely via navigang to the Home or
Manual menus.
3.3.2.Program Progress Menu
Once a channel is selected for a program via the Channel Selecon
menu, the Program Progress menu is displayed, as shown in Figure
10. The menu lists various informaon regarding the respecve
progress of the programs running in pressure channel A and
pressure channel B. It also contains controls for manually stepping
through cycle steps for each program. If only a single channel is
being run, the menu will display informaon about the pressure
setpoint and me duraon for the next step in the current program
instead of the live pressure reading from the unused pressure
channel.

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Figure 10: Annotated drawing of Program Progress menu with 2
programs running at once (1 for channel A, 1 for channel B).
1. Current program: the currently-running and currently-
controllable program is shown here. Depending on which set
of controls for which channel is currently being hovered over
(Figure 10 opons 6 and 7), the corresponding program for
that channel will be shown.
2. Step, esmated me remaining informaon: the current step
and total number of steps in the program are displayed along
with the esmated me remaining (in the format of
minutes:seconds)11. Ticking of the esmated me remaining
countdown is triggered whenever the setpoint pressure of
the current step is reached. If the program has completed its
run and is on the nal step, a message saying “COMPLETE!”
will be shown in place of a me remaining esmate.
3. Pressure, me setpoint informaon: the pressure setpoint
and me duraon for the current program step are
displayed12.
4. Current pressure for Channel A: a live readout of the pressure
output by Channel A is displayed here. A checkmark (and
stopwatch, indicang the mer clock is proceeding) symbol is
displayed whenever the setpoint pressure is achieved.
5. Current pressure for Channel B: same funconality as Current
pressure for Channel A but for channel B pressure.
6. Channel A controls: this buon array contains various
controls for manually navigang an automac program cycle
in channel A should that be necessary. For descripons of the
funcon of each control, see Figure 10 opons 8-11 in this
list. If only one program is selected and running, only a single
set of controls will be displayed corresponding to that chosen
program.
7. Channel B controls: same funconality as Channel A controls
but for channel B pressure.
8. Step back buon: opon to go back one step in the
currently running program.
9. Stop/ Programs buon: opon that pauses the program
and turns o output pressure, eecvely stopping the
program and any output of the pressure channel. When
Step Forward
buon (Figure 10 opon
11). More informaon on
these hold steps can be
found in the vivoChip-2x
manual (VVS-QA-015),
included with vivoChip-2x
device sets.

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/
stopped, the buon will be displayed, giving the user the
opon to substute a program from the Programs menu in
place of the stopped program.
10. Play/ Pause buon: opon that starts/resumes ( )
or pauses ( ) the selected program. Pausing the program
resets the setpoint mer, but it maintains the setpoint
pressure of the current step unl the program is resumed by
selecng the Play buon.
11. Step Forward buon: opon to proceed to the next
step in the running program. If the current step is a “hold”
step, it is required to select this opon to proceed within the
program.
When a program proceeds to its last step and achieves the setpoint
pressure and me duraon, the program is considered to be
complete. The pressure and me setpoint of the nal step in the
program will hold indenitely unl the user navigates out of the
Programs menu. As a general rule, leaving the Programs menu
(via Home or Manual menu opons) turns o the pump and
any currently running programs—stay on the Programs menu or
associated program status menu to ensure the compleon of any
selected programs.
An excepon to the preceding general rule occurs if and only if the
following condions are all met:
1. A program has reached compleon and reads “COMPLETE!”
for one or both pressure channels
2. The user navigates directly to the Manual menu from the
Programs menu
Under this circumstance, the Manual menu will start up with the
nal pressure seng for each complete channel already running.
This is useful if the user wishes to modulate the nal holding
pressure manually without losing applied pressure during menu
navigaon.
3.4. Sengs
The Sengs menu, depicted in Figure 11, contains various user-
changeable sengs that aect the operaon of the vivoCube+.

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/
Select a seng to change it using the mulfuncon dial, and select
once more to apply the seng. The vivoCube+ will retain these
sengs between power cycles.
Figure 11: Drawing of Sengs menu.
1. Pressure Source: this seng allows the user to choose
whether the vivoCube+ should use the built-in pressure
source (pump installed inside the vivoCube+) or an external
pressure source provided by the user connected to the
external pressure source port (Pext). For informaon on how
to use an external pressure source with the vivoCube+, see
Appendix A. The default seng is to use the built-in pressure
source.
2. Sound Alerts: this seng toggles the system sound alerts on
or o. A chime will play when the built-in pump turns on or
o, when a program commences, and when a program
completes when sound alerts are set to on. When set to o,
no alert sounds will play. The default seng is on.
3. LED Ring Brightness: this seng allows the user to set the
brightness of the LED ring indicator surrounding the mul-
funcon dial control on the front panel of the vivoCube+.
Brightness can be set from 0 to 100% in increments of 5%.
The default value is 100%.
4. Screen Brightness: this seng allows the user to set the
screen brightness from 20 to 100% in increments of 5%. The
default value is 100%.
5. Info: clicking this opon will display system informaon on
the Info menu (soware version and system serial