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Discover the Drivers of Cell Fate,
Function, and Fitness
Agilent Seahorse XF analysis platform
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Glucose
Cell
Respiration
H
+
O
2
H2O
Glycolysis
ATP
ADP
ATP
ADP
Are You Measuring What’s
Really Important to Your Cells?
To fully understand what drives cell phenotype and function, you must consider the
influences of energy metabolism.
Examining energy metabolism has led to new insights into biological function. In fact,
some of the decade’s most significant discoveries have hinged upon elucidating the role
of energy metabolism in cellular processes.
Live-cell, functional metabolic data is impacting research and discovery in areas like these:
Immunology
– Determining metabolic fitness of immune cells
– Measuring and modulating immune cell activation
in real time
– Targeting metabolic pathways to improve
immunotherapy design and performance
Cancer
– Rewiring metabolism to support rapid growth
– Understanding which nutrients fuel cancer cells
– Uncovering how metabolism impacts the tumor
microenvironment
Toxicology
– Detecting mitochondrial liabilities among
potential therapeutics
– Assessing the risk of mitochondrial toxicity in vitro
with high specificity and sensitivity
– Determining the mechanism of action of
mitochondrial toxicants
Obesity, diabetes, and metabolic disorders
– Measuring functional effects of genetic
changes to metabolic pathway components
– Examining nutrient use in healthy and
diseased cell models
– Assessing fatty acid oxidation and glycolysis
in different cell types
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Complete Platform for Live-Cell Metabolic Assays
Comprising instruments, assay kits, software, and consumables, Agilent Seahorse XF platforms
are designed to support you from sample to insight.
State-of-the-art
data analytics tools
Real-time
calculation
of results
Validated
kits, media,
and reagents
Label-free
pH and O
sensor cartridge
A
D
B
C
More relevant
injection ports
for real-time
modulation
2
Live-cell
analysis
with 2D and 3D
plate options
Move beyond analyzing what your cells are and get a clear picture of what they do
– Instrumentation for real-time extracellular flux (XF) measurements in live cells
– Validated kits, media, and reagents for standardized XF assays
– Patented sensors, plastics, and consumables for reliable XF data
– Powerful analytics to help interpret and share your results
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Agilent Seahorse XF analyzers: The standard for measuring energy metabolism in live cells
Seahorse XF analyzers enable robust measurements of mitochondrial activity, glycolysis, and ATP production
rates in a microplate format.
– Automatic calculation of oxygen consumption and proton efflux rates
– Label-free detection of discrete bioenergetic changes in live cells—in real time
– Compatibility with both adherent and suspension cells, as well as isolated mitochondria
– Instrument-controlled gentle mixing, measuring, and compound injections
– High sensitivity to analyze as few as 5,000 cells per well
– Temperature controlled to maintain cell health and kinetics
Agilent Seahorse XF HS
Mini Analyzer
High-sensitivity capabilities—combined
with an easy-to-use interface—make
this eight-well analyzer accessible for
every lab while delivering robust XF
analysis on limited cell numbers. It also
allows you to use fewer cells per well,
compared with other XF platforms.
Key applications:
– Optimization of experimental
conditions before scaling up
– Rare, sorted, and/or primary
immune cells
– Quiescent and low-respiring
cell types
– Cell lines and ex-vivo samples
Agilent Seahorse XFe96 Analyzer
Maximize your capacity for XF assays
and minimize cost per sample.
The Seahorse XFe96 enables many
experimental groups per assay,
maximizing design flexibility and
sample throughput.
Key applications:
– Phenotypic screening
– Testing many conditions
at one time
– Dose-response studies
– Spheroids
Agilent Seahorse XFe24 Analyzer
Featured in hundreds of peer-reviewed
publications, the Seahorse XFe24
uses a larger well format suitable for
islets and other specialty samples.
Key applications:
– Islets
– Small-model organisms
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Take a Closer Look at Seahorse XF Technology
The patented Agilent Seahorse XF platform lets you measure both metabolic energy pathways
simultaneously with speed and precision.
Integrated injection ports
sequentially deliver up to
four compounds, allowing
you to test multiple
conditions per well.
Cells are seeded in the wells
of the custom Seahorse XF
cell culture microplate.
Sensor probes gently lower to
create a transient microchamber.
This process allows rapid, real-time
measurement of changing oxygen
and proton concentrations in the
extracellular medium.
Biologically compatible
optical microsensors
simultaneously measure
rates of oxygen
consumption and
extracellular acidification,
without adding dyes.
The microplate
well requires 10- to
20-fold fewer cells
than conventional
respirometers.
Instrument mechanics
gently raise and lower the
cartridge to mix the fluid
in the wells for optimal
dispersal of modulators
and analytes.
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Quality XF sensor cartridges and cell plates are integral to the Seahorse XF measurement system
XF sensor cartridges: Essential to sensitive, simultaneous measurement
of bioenergetic pathways
– Non-invasive sensors measure dissolved oxygen and free protons.
– Built-in injection ports deliver compounds at preprogrammed intervals.
– Precision manufacturing process ensures consistent lot-to-lot performance.
XF cell culture microplates: Specific to application and instrument type
– Plates are tissue-culture treated and irradiated for cell culture operations.
– Unique well geometry enables rapid, sensitive XF measurements.
– Adheres to SBS standards for plate reader compatibility.
– Poly-D-Lysine coated plates ensure more consistent suspension cell workflows.
– XF HS miniplate accommodates fewer cells per well when sample amount is limited.
Did you know...
Having papers with functional data improves your chances of getting published
in a top-tier journal?
Observing intact cell behavior in real time provides much-needed context and confirmation of
omics data. No wonder it’s playing a growing role in peer-reviewed literature..
Nearly 7,000 publications to date
Agilent cell analysis publications (through 2019)
7,000
6,000
5,000
4,000
3,000
2,000
1,000
Annual
Cumulative
0
2009 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019
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Oxygen Consumption Rate (OCR)
Gain Deeper Insight into Your
Time (min)
Oxygen consumption rate (OCR)
0
Bioenergetic profile
ATP production rate (pmol/min)
Glycolytic rates
Cell Function with Agilent
Seahorse XF Assay Kits
How are the cells generating energy? Has my treatment changed the phenotype? Agilent Seahorse XF Assay kits—built on
validated biology—help you answer these questions and more.
Mitochondrial respiration
Oligomycin FCCP
360
320
280
240
200
160
(pmol/min)
Basal
120
respiration
80
40
Non-mitochondrial oxygen consumption
0
0 20 40 60 8010 30 50 70 90 11
50%
65%
50%
35%
Rot/AA 2-DG
400
Compensatory
300
200
Mito
acidification
100
Basal
glycolysis
Proton efflux rate (pmol/min)
0
0 10 20 30 40 50 60 70
Glycolytic function
Glucose Oligomycin 2-DG
45
40
35
30
25
20
(mpH/min)
15
Glycolysis
10
5
Extracellu lar acidification ra te (ECAR)
Non-glyc olytic acidif ication
0
0 20 40 60 8010 30 50 70 9 0 10
ATP-linked
respiration
Proton leak
glycolysis
25%
75%
Time (min)
Time (min)
Glycolytic
capacity
Maximal
respiration
Mito
Glyco
Rotenone and
Antimycin A
Total proton efflux
Glycolytic proton efflux
Spare
capacity
Total
Glycolyti c
reserve
Agilent Seahorse XF Cell Mito Stress Test Kit
This assay is well-recognized for gaining insight into the components of mitochondrial function.
Investigate functional differences among cell types and drug candidates, as well as genetic or
biochemical interventions.
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Agilent Seahorse XF Real-Time ATP Rate Assay Kit
The only assay that measures the rates of ATP production from glycolysis and mitochondrial
respiration simultaneously. Quantification of ATP production rate delivers a dynamic picture of
cellular bioenergetics, giving you a unique understanding of cellular phenotype and function.
Agilent Seahorse XF Glycolytic Rate Assay Kit
Precisely measures glycolysis in live cells, revealing transient responses and rapid metabolic
switches not discernible in endpoint lactate assays. You can also quantify proton efflux
specific to glycolysis.
Agilent Seahorse XF Substrate Oxidation Kits
A suite of optimized kits for rapidly measuring cellular substrate oxidation by assessing changes
in oxygen consumption (OCR) in live cells. These kits facilitate investigation of how cells alter
or shift oxidation of specific substrates to perform key cellular functions, as well as respond to
genetic or pharmaceutical interventions.
0
8
Aerobic
Metabolic Potential
Baseline
Mitochondrial Respiration
Phenotype
Quiescent
Glycolysis
Extracellular Acidification Rate (ECAR)
Energetic
Stressed
Phenotype
Glycolytic
Agilent Seahorse XF Cell Energy Phenotype Test Kit
This simple assay simultaneously measures mitochondrial respiration and glycolysis with just
a single injection, so you can rapidly determine energy phenotypes of cells and investigate
metabolic switching. It’s also a useful assay for screening compounds that can modulate
mitochondrial and/or glycolytic function.
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Transform Data into Insights
Control without the complexity
The Agilent Seahorse XFe Analyzer and XFp Analyzer
software interfaces provide an intuitive, streamlined
experimental setup and data acquisition experience.
– Predefined templates designed for each Seahorse XF
Assay Kit ensure accurate, consistent experiment setup.
– Kinetic rate data is automatically calculated and
displayed in real time, allowing you to observe timedependent biological responses throughout your assay.
Turn raw metabolic data into publishable results
Agilent Seahorse analytics, the next generation of XF data
analysis, provides secure data storage and intuitive data
analytics in the cloud. Features include:
– Desktop-like interactivity for accessibility from
anywhere on both PC and Mac
– Built-in formulas for automatic calculations and
summaries of Seahorse XF assay parameters
– Customizable analysis views for turning data to
insight—or for easy data sharing
– Flexible tools for easily exporting your data into third-
party graphing and statistical software applications,
such as GraphPad Prism and Microsoft Excel
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Complete Your Workflow—
and Advance Your Live-Cell
Metabolic Analysis
Agilent offers a complete workflow for live-cell metabolic analysis including cell metabolism analyzers, liquid handling
automation, imaging plate readers, and high-quality consumables. All work together to help you achieve consistent,
reproducible data.
Sample preparation
Seed cells
Manual or Bravo
Wash
cells
Bravo
Image and
incubate
Cytation imager with imaging
and normalization solution
Consistent data starts with consistent
sample preparation
Getting reliable data from live cell-based assays can be
challenging if your assay prep methods are not reliable.
Automated sample preparation can maximize productivity
and minimize human error.
The Bravo Seahorse Assay Workbench allows users of all
experience levels to:
– Standardize sample preparation. Intuitive software
ensures consistent results between users.
– Increase walk-away time, improve the efficiency of
your lab personnel.
– Minimize variability, generating consistent data no matter
who is running the protocol.
Dilute
compounds
Bravo
Load cartridge
ports
Bravo
Run assay
Seahorse XFe96 Analyzer
Analyze data
Wave software
Seahorse analytics
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Integrate imaging and normalization into your Seahorse XF analysis platform for
more interpretable results
The BioTek Cytation combines automated digital microscopy and conventional microplate detection
in a configurable, upgradable platform.
– Take advantage of powerful imaging, data capture, and data analysis.
– Count cellular objects in situ.
– Easily document wells of interest.
– Normalize cell count to compare across treatment groups.
– Seamlessly integrate normalization data and well images into your XF assay result file.
BioTek Cytation
Seahorse Wave controller software
Seahorse XFe Analyzer
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Agilent Value Promise
We guarantee you at least 10 years of instrument use from your
date of purchase. Otherwise, we will credit you with the residual
value of the system toward an upgraded model.
Learn more about Agilent Seahorse XF technology:
www.agilent.com/chem/discoverXF
Find a local Agilent customer center:
www.agilent.com/chem/contactus
U.S. and Canada
1-800-227-9770; select option 3, then 8
cellanalysis.support@agilent.com
Europe
UK: 0800 096 7632
Germany: 0800 180 66 78
Netherlands: 0800 022 7243
Other EU: +45 3136 9878
cellanalysis.support@agilent.com
Asia Pacific
cellanalysis.support@agilent.com
For Research Use Only. Not for use in diagnostic procedures.
RA.5694791667
This information is subject to change without notice.
© Agilent Technologies, Inc. 2021
Published in the USA, February 1, 2021
5994-1329EN
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