Agilent Technologies Seahorse XFe Manual de usuario

Seahorse XFe Analyzer

Operating Manual

Seahorse XFe Analyzer Operating Manual

Notices

No part of this manual may be repro-

duced in any form or by any means

(including electronic storage and

retrieval or translation into a foreign lan-

guage) without prior agreement and

written consent from Agilent Technolo-

gies, Inc. as governed by United States

and international copyright laws.

Manual Part Number

S7804-90001_REV A

Edition

Second Edition, June 2018

First edition, December 2012

Printed in USA

Agilent Technologies, Inc.

121 Hartwell Ave.

Lexington, MA 02466

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to being changed, without notice, in

future editions. Further, to the maxi-

mum extent permitted by applicable

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chantability and fitness for a particular

purpose. Agilent shall not be liable for

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Safety Notices

CAUTION

A CAUTION 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 damage to the product

or loss of important data. Do

not proceed beyond a CAUTION

notice until the indicated

conditions are fully understood

and met.

WARNING

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.

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