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
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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
death. Do not proceed beyond a
WARNING notice until the
indicated conditions are fully
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.