thE world’S moSt AdvAncEd mEtABolic AnAlyzErS

Seahorse XFe Extracellular Flux Analyzers

cancer immunology Stem cell Biologycardiovascular disease neurodegeneration toxicology and hepatobiologycell physiology obesity, diabetes, and metabolic disorders translational medicine

the role of metabolism in cellular and physiological processes, from cardiovascular function to immune

response, is well established; many diseases are now linked to metabolic dysfunction or reprogramming.

using the Seahorse XFe Extracellular Flux Analyzer, scientists are now able to easily obtain powerful

functional metabolic data to gain a greater understanding of cell metabolism and enable new

advancements in life science research.

Functional Metabolic Data on Live Cells in Real Time

the Seahorse XF cell Energy phenotype test measures the key parameters used to determine the energy phenotype of a cell: baseline phenotype, stressed phenotype, and metabolic potential.

the Seahorse XF glycolysis Stress test reports three key parameters of glycolytic function: glycolysis, glycolytic capacity, and glycolytic reserve.

the Seahorse XF Fatty Acid oxidation assay measures both exogeneous palmitate oxidation and endogenous fatty acid oxidation.

the Seahorse XF cell mito Stress test measures the key parameters of mitochondrial function: basal respiration, Atp production, proton leak, maximal respiration, and spare respiratory capacity.

Extracellular Acidification Rate (ECAR)

Oxy

gen

Con

sum

ptio

n R

ate

(OC

R)

Glycolysis

XFp Cell Energy Phenotype Profile

Mito

chon

dria

l Res

pira

tion

Aerobic

Quiescent

Energetic

Glycolytic

BaselinePhenotype

StressedPhenotype

Metabolic Potentia

l

Oxy

gen

Con

sum

ptio

n R

ate

(OC

R)

Seahorse XF Fatty Acid Oxidation AssayExogenous & Endogenous Fatty Acid oxidation

Oxy

gen

Con

sum

ptio

n R

ate

(OC

R)

Oxy

gen

Con

sum

ptio

n R

ate

(OC

R)

Seahorse XF Glycolysis Stress Testglycolytic Function

Seahorse XF Cell Mito Stress Testmitochondrial respiration

TIME (minutes)

(mpH

/min

)

Glycolytic Function

Glucose Oligomycin 2-DG

0

5

10

20

30

15

25

35

40

45

0 20 40 60 8010 30 50 70 90 100

Non-glycolytic Acidification

Glycolysis

GlycolyticCapacity

GlycolyticReserve

(pm

ol/m

in)

TIME (minutes)

180

160

140

120

100

80

60

40

20

00 10 20 30 40 50 60

Rotenone & antimycin A

Oligomycin FCCP

ExogenousPalmitate Oxidation

ExogenousPalmitate Oxidation

Endogenous FAO

Basal

Maximal

XF Fatty Acid Oxidation Profile

Endogenous FAO

TIME (minutes)

(pm

ol/m

in)

Mitochondrial Respiration

Oligomycin FCCP Rotenone & antimycin A

0

120

80

40

160

200

240

280

320

360

0 20 40 60 8010 30 50 70 90 100 110

Non-mitochondrial Oxygen Consumption

BasalRespiration

ATPProduction

Proton Leak

MaximalRespiration

SpareCapacity

Extr

acel

lula

r Aci

dific

atio

n R

ate

(EC

AR

)

Extracellular Acidification Rate (ECAR)

SeahorSe XF aSSayS — The Gold STandard For MeTabolic MeaSureMenTS

Seahorse XF Cell Energy Phenotype Testmetabolic phenotype & potential

CancerSeahorse XF glycolysis Stress test reveals mechanism of action of cancer pathway inhibition.

Obesity/DiabetesSeahorse XF technology reveals hedgehog pathway-induced switch to a glycolytic phenotype in adipocytes.

NeurodegenerationSeahorse XF cell mito Stress test reveals metabolic signatures of embryonic stem cells with huntington mutations.

ImmunologySeahorse XF cell mito Stress test demonstrates fatty acid oxidation is required for memory t cell formation.

viale et al., (2014) oncogene ablation-resistant pancreatic cancer cells depend on mitochondrial function. nature. 514(7524):628-32.

50

P < 0.01

4540353025201510

5

5 10 15 20 25 30 35 40 45 50 55 600

Control

PI3K/mTOR&MEK1PathwayInhibitors

GlucoseOligomycin

2DG

Aerobic

Glycolytic

MoreMetabolic

LessMetabolic

160

Control

SAG

140

120

1004 5 6 7

800

Control orEtomoxir Oligomycin

IL-15 + controlIL-15 + etomoxir

FCCPRotenone/

antimycin A

400

600

200

10025 50 75 100 120

Functional metabolic data is required to tell a complete story of cellular processes and pathologies.

the Seahorse XFe Extracellular Flux Analyzer provides the capability to examine the intact and functional

cellular system, enabling comprehensive investigations into metabolic pathways, substrate preference and

utilization, catabolic and anabolic processes, and metabolic phenotypes.

SeahorSe XF TechnoloGy — GeneraTinG new inSiGhTS inTo diSeaSeo

xyge

n c

onsu

mpt

ion

rat

e (o

cr

)(p

mol

/min

)

oxy

gen

con

sum

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n r

ate

(oc

r)

(pm

ol/m

in)

oxy

gen

con

sum

ptio

n r

ate

(oc

r)

(pm

ol/m

in)

Extracellular Acidification Rate (ECAR) (mph/min)

Ext

race

llula

r Aci

dific

atio

n R

ate

(EC

AR

) (m

ph/m

in)

time (min) time (min)

time (min)

600

500

Oligomycin

Embryonic lethal mutationControl

FCCP Rotenone/antimycin A

300

400

200

100

0100 20 30 40 50 60 70

Dominantly inherited Huntington’s Disease

human pancreatic tumor cells human pancreatic tumor cells

murine adipocytes murine embryonic stem cells

van der windt et al., (2012) mitochondrial respiratory capacity is a critical regulator of cd8+ t cell memory development. immunity. 36(1): 68-78.

teperino et al., (2012) hedgehog partial agonism drives warburg-like metabolism in muscle and brown fat. cell. 151(2): 414-26.

ismailoglu et al., (2014). huntingtin protein is essential for mitochondrial metabolism, bioenergetics and structure in murine embryonic stem cells. dev. Biol. 391(2): 230-40.

How XF Technology Works

the Seahorse XFe Analyzer simultaneously measures the two major energy pathways of the cell — mitochondrial respiration and glycolysis — in live cells using a label-free, solid-state sensor cartridges in a microplate format. the Seahorse XFe Analyzer works with all cell types, including primary cells, cell lines, suspension cells, as well as islets, C. elegans, yeast, and isolated mitochondria.

thE pAtEntEd microchAmBEr mAkES it All poSSiBlE

A

B

C

D

E

integrated injection ports sequentially deliver up to 4 compounds allowing dose response, agonist or antagonist response, or pathway perturbation analysis of each sample.

Sensor probes gently lower to create a transient microchamber, allowing rapid, real time measurement of changes in both oxygen and proton concentrations in the extracellular medium.

Seahorse XF cell culture microplates support virtually all cell types.

inert optical micro sensors measure rates of oxygen consumption and extracellular acidification simultaneously.

the microplate well requires a small number of cells, 10-20 fold fewer cells compared to conventional respirometers.

I

II

III

IV

Lactate

Pyruvate

Glucose

GLYCOLYSIS — ECAR (Extracellular Acidification Rate)

cells generate Atp via glycolysis independent of oxygen, producing lactic acid and protons.

the Seahorse XFe Analyzer measures glycolysis by measuring the extracellular acidification rate (ECAR) of cells.

MITOCHONDRIAL RESPIRATION —

OCR (Oxygen Consumption Rate)

mitochondria consume oxygen when oxidizing fatty

acids or other substrates to generate Atp. the Seahorse

XFe Analyzer measures mitochondrial respiration by measuring the oxygen

consumption rate (ocr) of cells.

the Seahorse XFe Analyzer utilizes patented transient microchambers which enable sensitive, precise, and nondestructive metabolic measurements in real time.

Cutaway graphic of a single probe

and well

BC

E

D

A A

Seahorse XF Data in Publications there are over 2,000 papers utilizing Seahorse XF technology published in leading scientific journals such as nature, cell, Science and pnAS. Scientists are embracing Seahorse XF technology to identify metabolic phenotypes and reprogramming, as well as target metabolic changes for therapeutic purposes.

Built on innovative and proven Seahorse XF technology, the Seahorse XFe Extracellular Flux Analyzer

makes it easy to perform functional metabolic measurements in live cells, in real time. A complete platform

that includes hardware, software, consumables, and reagents, the Seahorse XFe Analyzer is a powerful

and flexible system that is accelerating life science research around the world.

complEtE plAtForm For mEtABolic ASSAyS

powErFul AnAlyticS

EASy-to-uSE kitS And

rEAgEntS

FunctionAl livE cEll ASSAyS

provEn, EnABling

tEchnology

EXpErt trAining And

Support

NEW

DISCOVERIES

© 2016 All rights reserved. Seahorse Bioscience and the Seahorse logo are trademarks of Seahorse Bioscience inc. All other trademarks and registered trademarks are the property of their respective owners. the content in this brochure is for informational purposes only and may be subject to change without notice.

Bringing a new perspective to cEll mEtABoliSm rESEArch

Specifications Seahorse XFe24 Analyzer Seahorse XFe96 Analyzer

Dimensions 41cm x 61cm x 43cm(16" x 24" x 17" )

41cm x 61cm x 43cm(16" x 24" x 17" )

Weight 20 kg (45 lb) 14 kg (35 lb)

Power Requirements 100-240vAc50/60hz

100-240vAc50/60hz

Primary Measurements Made ocr & EcAr ocr & EcAr

Assay Format live cells in a specialized 24-well microplate

live cells in a specialized 96-well microplate

Software

wave supports assay design, instrument control, and data analysis. can be installed on windows® pc for analysis or data export to mS® Excel™ or graphpad prism®.

wave supports assay design, instrument control, and data analysis. can be installed on windows® pc for analysis or data export to mS® Excel™ or graphpad prism®.

Injection Ports 4 per well, 75 µl each 4 per well, 25 µl each

Assay Running Volume 500-1000 µl/well 80-200 µl/well

Sample Requirements 10,000-1,000,000 cells/well 4,000-500,000 cells/well

Instrument Controllercombination computer and touch-screen display with full assay design, control, and analysis capability.

combination computer and touch-screen display with full assay design, control, and analysis capability.

Best For/Key Advantages islets

Maximum experimental flexibilityhighest throughput Spheroids dose-response studies

cancer immunology Stem cell Biologycardiovascular disease neurodegeneration toxicology and hepatobiologycell physiology obesity, diabetes, and metabolic disorders translational medicine

rev

3

Corporate Headquarters

Seahorse Bioscience inc.16 Esquire roadnorth Billerica, mA 01862 uSphone: +1 978 671 1600www.seahorsebio.com

European Headquarters

Seahorse Bioscience EuropeFruebjergvej 32100 copenhagen dkphone: +45 31 36 98 78

Asia-Pacific Headquarters

Seahorse Bioscience Asia199 guo Shou Jing rd, Suite 207pudong, Shanghai 201203 cnphone: 0086 21 33901768