Agilent Technologies Seahorse XFe User manual

Seahorse XFe Analyzer
Operating Manual

Seahorse XFe Analyzer Operating Manual
Notices
© Agilent Technologies, Inc. 2018
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Manual Part Number
S7804-90001_REV A
Edition
Second Edition, June 2018
First edition, December 2012
Printed in USA
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hazard. It calls attention to an
operating procedure, practice,
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A WARNING notice denotes a
hazard. It calls attention to an
operating procedure, practice,
or the like that, if not correctly
performed or adhered to, could
result in personal injury or
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understood and met.

Seahorse XFe Analyzer Operating Manual 3
About This Manual
This manual contains information for operating and maintaining the Agilent
Seahorse XFe Analyzer.
1“Introduction”
Chapter 1 provides an introduction to the Agilent Seahorse XFe Analyzer.
2 “Installation”
Chapter 2 provides unpacking and installation instructions for the Agilent
Seahorse XFe Analyzer.
3“BasicOperation”
Chapter 3 provides basic operating procedures for the Agilent Seahorse XFe
Analyzer.
4 “Maintenance”
Chapter 4 provides routine maintenance, troubleshooting, contact, and additional
resource information for the Agilent Seahorse XFe Analyzer.

4 Seahorse XFe Analyzer Operating Manual

Seahorse XFe Analyzer Operating Manual 5
Contents
About This Manual 3
1Introduction
General Information 8
Safety Considerations 9
Electrical hazards 9
Instrument Overview and Intended Use 10
Technical Specifications 11
2 Installation
Unpacking and Component Identification 14
Installation Procedure 17
Suitable locations for the XFe system 17
Internal components of the XFe Analyzer 18
Setup and interconnects: cable installation 20
3 Basic Operation
Power and Warm Up 24
Power up 24
Launch Wave 24
Performing XF assays 25
XFe status indicator 25
Wave Controller widgets 26
XFe Assays at Non-37 °C Temperatures 28
Operational and assay guidelines for non-37 °C
assays 29
Set alarm (temperature tolerance range) 30
4 Maintenance
Cleaning and Routine Maintenance 32

6 Seahorse XFe Analyzer Operating Manual
Troubleshooting 33
Barcode errors 33
Contact Information 36
Worldwide technical support 36
Ordering 36
Additional Resources: 37

Seahorse XFe Analyzer Operating Manual 7
1Introduction
General Information 8
Safety Considerations 9
Electrical hazards 9
Instrument Overview and Intended Use 10
Technical Specifications 11
This chapter provides an introduction to the Agilent Seahorse XFe Analyzer.

1Introduction
General Information
8 Seahorse XFe Analyzer Operating Manual
General Information
This manual covers several models of Agilent Seahorse XFe Analyzer:
Instrument type Model numbers (32-bit) Model numbers (64-bit)
Seahorse XFe96 101991-100, S7800A S7800B
Seahorse XFe24 102238-100, S7801A S7801B

1Introduction
Safety Considerations
Seahorse XFe Analyzer Operating Manual 9
Safety Considerations
The XFe has been carefully designed so that when used properly you have an
accurate, fast, flexible, and safe instrument.
Information on safety practices is provided with your instrument and operation
manuals. Before using your instrument or accessories, you must thoroughly read
these safety practices.
Electrical hazards
Panels or covers that are retained by fasteners which require the use of a tool for
removal may be opened only by Agilent-trained, Agilent-qualified, or
Agilent-authorized service engineers. Consult the manuals or product labels
supplied with the XFe to determine which parts are operator-accessible.
Always use a three-wire outlet with ground connection which is adequately rated
for the load. The installation must comply with local safety regulations.
WARNING
Observe all relevant safety practices at all times.
If the equipment is used in a manner not specified by the manufacturer, the
protection provided by the equipment may be impaired. Unskilled, improper, or
careless use of this instrument can create shock hazards, fire hazards, or
other hazards which can cause death, serious injury to personnel, or severe
damage to equipment and property.
WARNING
The XFe contains electrical circuits, devices, and components operating at
dangerous voltages. Contact with these circuits, devices, and components
can cause death, serious injury, or painful electric shock.
WARNING
Connection of the instrument to an incorrectly wired supply outlet, or lack of
proper electrical grounding can create a fire hazard or a potentially serious
shock hazard and could seriously damage the instrument and any attached
ancillary equipment.

1Introduction
Instrument Overview and Intended Use
10 Seahorse XFe Analyzer Operating Manual
Instrument Overview and Intended Use
The Agilent Seahorse XFe analyzer measures the rate of change of dissolved
oxygen and pH in the media immediately surrounding living cells cultured in a
microplate. Changes in the extracellular media are caused by the consumption
or production of analytes by the cells. Therefore, a sensitive measurement of the
media flux can be used to determine rates of cellular metabolism with great
sensitivity and in a totally non-invasive, label-free manner.
A unique feature of the Agilent Seahorse XF technology is its ability to make
accurate and repeatable measurements in as little as five minutes. The
instrument, working with a sensor cartridge, isolates a few µL of media above the
cell monolayer. Cellular metabolism causes rapid, easily measured changes to
the “microenvironment” in this small volume.
An XFe sensor cartridge is required to run an assay. The cartridge has 24 or 96
probes, and each probe has a single multifluor sensor spot that is sensitive to
both oxygen and proton concentration. The system measures the concentration
of each analyte over time and automatically calculates the oxygen consumption
rate (OCR) and extracellular acidification rate (ECAR) simultaneously in every
well of the microplate.
Typically, a measurement cycle is performed for 5-8 minutes. The media is
gently mixed, the probe is positioned 200 µm above the well bottom, and the
analyte levels are measured until the oxygen concentration drops approximately
20-30% and media pH declines approximately 0.1-0.2 pH units.
Baseline metabolic rates are typically measured 3-4 times, and are reported in
pmol/min for OCR and in mpH/min for ECAR, compound is added to the media
and mixed, and then the post-treatment OCR and ECAR measurements are
made and repeated. As cells shift metabolic pathways, the relationship between
OCR and ECAR changes.
The XFe system, comprising a bench top analyzer and touch screen controller, is
driven by Wave software. This software enables all aspects of Seahorse assays
including assay setup, instrument control, and data analysis.
Consumables are sold separately and include FluxPaks (comprising sensor
cartridges, cell plates, and calibrant) as well as a variety of Assay Kits, Reagents,
and Media. XFe Sensor Cartridges are specific for the instrument type and can
be purchased exclusively from Agilent.

1Introduction
Technical Specifications
Seahorse XFe Analyzer Operating Manual 11
Technical Specifications
Model number 101991-100, S7800A (XFe96)
102238-100, S7801A (XFe24)
S7800B (XFe96)
S7801B (XFe24)
Part number 101991-100 (XFe96)
102238-100 (XFe24)
Dimensions
Controller:
Analyzer:
Width × height × depth
19" × 17" × 12"
48 cm × 43 cm × 30 cm
16" × 24" × 17"
41 cm × 61 cm × 43 cm
Weight Analyzer: 45 lbs/20 kg
Controller: 22 lbs/9 kg
Power requirements 100-240 V AC 50/60Hz
Analyzer: 6 A
Controller: 3.2 A
Power cord rating 3- wire (grounded) AC power cord rated 10 A or greater
Power fuse ratings 250 V/10 A Time delay fuses
Environmental conditions "Normal" Environmental conditions- indoor use, altitude to 2,000 m
Room temperature range +40 °F - 86 °F (+4 °C - 30 °C)
No direct sunlight
Humidity less than 80%
Sample temperature and
environment
Controlled to user-selected temperature between 16 °C and 42 °C,
but at least 12 °C above ambient temp
No gas or humidity control
Software OS Windows 7, 32 bit Windows 10, 64 bit
Data interface RS232c (internal)
TCP/IP (external)
USB Type B
64 bit barcode reader

1Introduction
Technical Specifications
12 Seahorse XFe Analyzer Operating Manual

Seahorse XFe Analyzer Operating Manual 13
2Installation
Unpacking and Component Identification 14
Installation Procedure 17
Suitable locations for the XFe system 17
Internal components of the XFe Analyzer 18
Setup and interconnects: cable installation 20
This chapter provides unpacking and installation instructions for the Agilent
Seahorse XFe Analyzer.

2Installation
Unpacking and Component Identification
14 Seahorse XFe Analyzer Operating Manual
Unpacking and Component Identification
The XFe Analyzer system is packaged in two boxes.
Upon receipt, immediately check each box for damage. Shipping damage must
be reported to the transportation company and Agilent. See “Contact
Information” on page 36.
The analyzer is shipped with protection components that must be removed prior
to use. Agilent recommends these components be removed by Agilent personnel
during installation.
To prevent damage during shipping, the instrument is shipped with a cartridge
loaded onto the probe head and lowered onto a plate on the tray. These items
must be removed prior to running the first assay. Agilent personnel will remove
these shipping protection components from the XFe Analyzer During Installation.
WARNING
The XFe Analyzer requires two people to lift and handle safely.
Each person should firmly grasp the base of the unit at opposite ends to each
other. Use OSHA standards for lifting techniques.
CAUTION
XFe Instruments must be installed by trained Agilent personnel ONLY.
Instrument/Component Quantity Image
XFe Instrument 1

2Installation
Unpacking and Component Identification
Seahorse XFe Analyzer Operating Manual 15
XFe Controller 32 bit
OR
XFe Controller 64 bit
1
1
Power cord (instrument) 1
Power cord (controller) 1
Power supply (controller) 1
RS232 cable 1
Instrument/Component Quantity Image

2Installation
Unpacking and Component Identification
16 Seahorse XFe Analyzer Operating Manual
USB cable 1
USB extension cable 1
Extra fan filters 2
Instrument/Component Quantity Image

2Installation
Installation Procedure
Seahorse XFe Analyzer Operating Manual 17
Installation Procedure
The following items are included in an XFe system:
•XFeAnalyzer- The analyzer is a temperature controlled instrument
containing all optical and electronic measurement components to measure
oxygen and proton flux of cells grown in XF cell culture plates. The analyzer is
used in conjunction with XFe sensor cartridges.
• Controller - Operation of the Analyzer is done through a high-resolution color
LCD touch screen with stand, that may be installed in front of or beside the
XFe. The controller communicates with the analyzer using a single serial
cable and a single USB A-B cable.
Suitable locations for the XFe system
XFe Analyzers are designed for laboratory use. The internal environment of the
analyzer is controlled to a preset temperature by the user; therefore, laboratory
room temperature must be maintained within the range listed in the specification
table.
Tray temperature control performance can be monitored using the status display
on the right side of the analyzer.
The XFe uses optical detection technology to measure extremely low levels of
fluorescent emission from analyte sensors. While the analyzer has been
designed to shield room light, excessive light (such as direct sunlight) should be
avoided.
WARNING
The electrical connection at the back of the XFe is the primary disconnect for
the instrument. The XFe should be positioned to allow accessibility to the
power cord for easy disconnection.
CAUTION
Avoid drafty areas, as well as areas experiencing significant vibration (such as
a centrifuge).

2Installation
Internal components of the XFe Analyzer
18 Seahorse XFe Analyzer Operating Manual
Internal components of the XFe Analyzer
Removing the side doors reveals the measurement chamber in which the assay
is conducted. The electro-optics hardware is enclosed in a card cage in the rear
chamber, and this is connected to the probe head through a set of fiber optic
cable bundles. The base of the enclosure contains the primary controller board
and heater assembly. (See Figure 1 and Figure 2.)
• LCD Message screen - Displays current instrument action and Thermal tray
temperature.
•Coverlatches- Pull on indented hand-holds molded into the side doors (not
shown) to lift them up, exposing the internal components of the instrument.
• Probe head and injection manifold - The probe head consists of 24 or 96
“light pipes” to carry the optical signals to and from the sensors. The Injection
manifold uses compressed air to inject compounds loaded into Sensor
cartridge ports into the assay wells.
Figure 1. XFe Front/Side view. Base color may vary.
LCD Message
screen
Cover latches (not
shown)
Fiber bundles
Thermal tray
Probe head and
Injection
manifold

2Installation
Internal components of the XFe Analyzer
Seahorse XFe Analyzer Operating Manual 19
•Frontstripperarm- Used for Sensor Cartridge removal at the end of assay
runs.
•Barcodereader- Reads bar code on sensor cartridge and cell plate.
• Circulating fan - Helps to stabilize environment inside XFe Analyzer.
Figure 2. XFe Front view
Front stripper
arm
Bar code reader
Circulating fan

2Installation
Setup and interconnects: cable installation
20 Seahorse XFe Analyzer Operating Manual
Setup and interconnects: cable installation
The XFe Analyzer is operated from a touch screen computer monitor mounted to
a stand, referred to as a controller. One RS232 cable and one USB cable handle
the communication of commands and data between the instrument and the
controller.
The controller may be connected to an external network, through the ports on the
underside.
See “Unpacking and Component Identification” on page 14 to identify each
cord, and refer to the figures below to identify the connectors.
1Connect power cords: One power cord is used to connect the instrument to a
grounded AC (mains) outlet. A second power cord is used to connect the
controller power supply module to the AC supply. The power supply module
is then connected to the socket at the bottom of the controller.
(See Figure 3.)
2Connect the data cables to the analyzer. One RS232 cable connects the
controller serial port to the analyzer socket labeled "COM." (See Figure 4.)
3A second cable (USB) connects the analyzer socket labeled "USB" to the USB
port on the controller directly adjacent to the network (Ethernet) port. This
port must be used for proper functioning of the instrument and barcode
reader. (See Figure 3.)
Figure 3. Controller ports (underside)
Figure 4. XFe rear panel - USB and serial ports
Serial port
Network port
Power input
USB ports (4)
Serial port for
controller
connection
USB ports from
barcode reader