Mar 11, 2019 the seahorse xf cell energy phenotype test in conjunction with a seahorse bioscience xf96 extracellular flux analyzer measures the mitochondrial respiration and glycolysis under baseline and. The xf96well microplates were analyzed using the seahorse bioscience xfe96 extracellular flux analyzer. Ocr and ecar measurements were performed using the xf24 or xf96 extracellular flux analyzer seahorse bioscience, north billerica, ma as described. Easily create assay protocols and analyze data with seahorse wave software.
Technologies seahorse bioscience is a leader in life sciences, diagnostics, and applied chemical markets. Furthermore, changes in mitochondrial function may regulate lymphocyte function in inflammatory diseases like type 2 diabetes. Extracellular flux analysis to monitor glycolytic rates. Seahorse bioscience xf96 extracellular flux analyzer. Seahorse xf analyzers measure oxygen consumption rate ocr and extracellular acidification rate. Box of seahorse xf96 polystyrene cell culture microplates pn 101085004. Livecell metabolic assay platform for life science research. Luz nicholas school of the environment, duke university, durham, north carolina.
Seahorse xfe96 extracellular flux analyzer the seahorse enables quantitation of metabolomics of live cells in real time with stimulation with up to 4 drugs. The new seahorse xfp analyser has been built on the basis of the proven. Agilent technologies seahorse bioscience technologies seahorse bioscience is a leader in life sciences, diagnostics, and applied chemical markets. The extracellular flux analyzer is a commonly used device for evaluating the performance of glycolysis and mitochondrial respiration in many cell types. Turn on instrument and start xf software to allow instrument to stabilize at 37 c. Nicholas school of the environment, duke university, durham, north carolina. Here, using the seahorse xf e 24 extracellular flux analyzer seahorse bioscience, massachusetts, usa we describe how to measure the fundamental parameters of the electron transport chain etc. Extracellular flux analyzer and prep station brand. This fast and sensitive measurement of cellular bioenergetics is label free, enabling timeresolved analysis and. Changes in the extracellular media are caused by the consumption or production of analytes by the cells. Ocr and ecar rates are key indicators of mitochondrial respiration and glycolysis and these measurements provide a systemslevel view of cellular metabolic function in. Xfe24 assays apply to all fields of biomedical research. Seahorse xfe96 analyzers measure ocr and ecar of live cells in a 96well plate format.
Agilent technologies seahorse bioscience in vitro technologies. Xf instruments measure cellular bioenergies in real time, in a microplate, and are used for scientific research, as well as drug discovery and development. Lowdose 2deoxyglucose and metformin synergically inhibit. The seahorse bioscience xf96 extracellular flux analyzer is an instrument that simultaneously measures two energy yielding pathways.
Seahorse xfp extracellular flux analyzer the role of metabolism in cellular and physiological processes from cardiovascular function to immune response is well established, with many diseases now linked to metabolic dysfunction or reprogramming. Seahorse xf96 installation and operation manual pdf. Seahorse bioscience bought bioprocessors corp in 2009. The agilent cell analysis software portfolio provides easytolearn, flexible toolssolutions for designing assays, instrument operation, analyzing data, and. High throughput screening of cellular bioenergetics.
Based analysis of cellular respiration in caenorhabditis elegans anthony l. The xf24 can also measure respiration of isolated mitochondria. Seahorse bioscience xfe instrument overview and intended use the seahorse bioscience xfe analyzer measures the rate of change of dissolved oxygen and ph in the media immediately surrounding living cells cultured in a microplate. The seahorse xf cell energy phenotype test in conjunction with a seahorse bioscience xf96 extracellular flux analyzer measures the mitochondrial respiration and. Accurate xf24 measurements require the analyzed cells to be grown in a. The seahorse bioscience xf96 extracellular flux analyzer figure 1 offers the advantage of significantly increased sensitivity, enabling the detection of defects in the oxphos system in human. Seahorse bioscience introduces xfe the worlds most advanced. This video demonstrates how the viaflo 96 handheld electronic pipette, used in conjunction with the seahorse bioscience xf96 extracellular flux analyzer, provides high throughput screening of the cellular bioenergetics of multiple cell lines. Seahorse technology is based on measuring the rate of extracellular fluxes of. Seahorse bioscience introduced extracellular flux technology to the life sciences market in 2006. Basal ecar were measured at three time points, followed by sequential injections of the glucose 10 mm, oligomycin 1. The extracellular flux xf assay described here is a platebased method in which galactoseconditioned hepg2 cells were acutely exposed. Seahorse bioscience introduces xfe the worlds most.
Integra has published a video demonstrating how its viaflo 96 handheld electronic pipette, used in conjunction with the seahorse bioscience xf96 extracellular flux analyzer, provides high throughput screening of the cellular bioenergetics of multiple cell lines. The xf96 extracellular flux analyzer seahorse bioscience is a noninvasive, labelfree, highthroughput instrument that measures the metabolic activity of living cells by simultaneously monitoring mitochondrial respiration and glycolysis. Seahorse xf96 v3 ps cell culture microplates, pack of 10. Rapid analysis of glycolytic and oxidative substrate flux. Extracellular flux analysis to monitor glycolytic rates and. The measurement of cellular bioenergetics, the processes by which cells produce and consume energy, is fundamental to determining. Launched in 2006, the awardwinning xf extracellular flux analyzer is the industry standard for measuring cellular bioenergetics, simultaneously measuring the two major energyproducing pathways. The oxygen consumption rate ocr and the extracellular acidification rate ecar were measured with a xf96 extracellular flux analyzer seahorse bioscience, agilent technologies.
The xfe24 flux analyzer measures real time kinetic of the media flux in live cellular environment to determine in vitro oxygen consumption rate ocr, and extracellular acidification rate ecar, in order to assess cellular metabolic functions such as oxidative phosphorylation, glycolysis, and fatty acid oxidation. Seahorse xf96 installation and operation manual pdf download. Its novel 8well miniplate format is ideal for use in pairwise comparison assays, as well as for use with primary cells and other precious samples. This instrument measures oxygen consumption rate ocr and extracellular acidification rate ecar of living cells simultaneously in a 24 well format.
Extracellular flux analysis to monitor glycolytic rates and mitochondrial oxygen consumption article in methods in enzymology 542. Measuring energy metabolism in cultured cells, including. Seahorse xf24 extracellular flux analyzer department of. Pdf the acute extracellular flux xf assay to assess. Good data with the software makes good graphic presentations. Only differences between the control group and the two hyperglycemic groups obp vs ogdm and obp vs ot1dm were analyzed. The company provides laboratories worldwide with instruments, services, consumables, applications, and expertise, enabling customers to gain the insights they seek. Based analysis of cellular respiration in caenorhabditis elegans. The seahorse bioscience xf96 system includes xf96 analyzer. The xf96 and xfe96 extracellular flux analyzer are 96 well tissue culture microplate systems allowing for a larger number of assays. The extracellular acidification rate ecar of niclosamide treated skov3 and ho8910 cells under the indicated conditions were measured in real time using the seahorse xf96 extracellular flux analyzer.
The extracellular flux xf assay described here is a platebased method in which galactoseconditioned hepg2 cells were acutely exposed to test compounds, then realtime changes in the oxygen consumption rate and extracellular acidification rate were simultaneously measured using a seahorse bioscience xf 96 analyzer. The seahorse extracellular flux xf analyzer seahorse bioscience is a. The agilent cell analysis software portfolio provides easytolearn, flexible tools solutions for designing assays, instrument operation, analyzing data, and. The seahorse extracellular flux xf analyzer seahorse bioscience is a powerful tool for measuring glycolysis and oxidative phosphorylation through oxygen consumption simultaneously in the same cells.
The seahorse bioscience xf e extracellular flux analyzer measures the two major energy pathways of the cell mitochondrial respiration and glycolysis in a microplate, in realtime. Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells. View online or download seahorse xf96 installation and operation manual. Seahorse xf96 manuals manuals and user guides for seahorse xf96. View and download seahorse xf96 installation and operation manual online. The seahorse xf24 is a revolutionary instrument for studies of cellular energy metabolism. Seahorse xf cell energy phenotype test kit receives top 10. Seahorse xfe96 extracellular flux analyzer dartlab. Software download for seahorse xf96 analyzer agilent. Quick overview preowned and in working condition computer included with software flux analyzer part no 100900101 pre station part no 101200100 manual included. Quick overview preowned and in working condition computer included with software flux analyzer part no 100900101 pre station part no 101200100 manual. Advances in the quantification of mitochondrial function.
This system has been used to study immune cells in a few published reports, yet a comprehensive protocol optimized particularly for lymphocytes is lacking. Rapid analysis of glycolytic and oxidative substrate flux of. The company provides laboratories worldwide with instruments, services, consumables, applications, and expertise. Analysis of lymphocyte mitochondrial function has been facilitated by introduction of 96well format extracellular flux xf96 analyzers, but the technology remains imperfect for analysis of human lymphocytes. Seahorse xfe24 extracellular flux analyzerbased analysis. Compact and easy to use, the xfp analyzer is designed to perform standard xf assays, and is ideal for pairwise comparisons. Novel technology opens the door to the understanding of. Xfe24 extracellular flux analyzer seahorse bioscience. The first published seahorse data was reported in a 2006 nature article. Seahorse xfe96 analyzer measures ocr and ecar of live cells in a 96well plate. Seahorse xfe analyzers from agilent cell analysis selectscience.
Seahorse wave desktop software is the assay design and data analysis software. The agilent seahorse extracellular flux analyzer can be used to measure the oxygen consumption rate ocr and extra cellular acidification rate ecar, from which mitochondrial bioenergetics measurements can be determined including basal respiration, respiration due to atp turnover, uncoupled respirationproton leak, and maximum respiration. Measurement of mitochondrial respiration and glycolysis allows energetic phenotyping of cells, in order to assess functional changes. The acute extracellular flux xf assay to assess compound. Seahorse bioscience xfe instrument overview and intended use the seahorse bioscience xfe analyzer measures the rate of change of dissolved oxygen and ph in the media immediately surrounding living cells cultured in a. Glycolysis is determined through measurements of the extracellular acidification rate ecar of the surrounding media, which is predominately from the excretion of lactic acid per unit time. Fetal hyperglycemia changes human preadipocyte function in. Seahorse bioscience xf96 extracellular flux analyzer product. Seahorse xf24 extracellular flux analyzer the seahorse xf24 is a revolutionary instrument for studies of cellular energy metabolism.
Advances in the quantification of mitochondrial function in. Seahorse bioscience xf96 extracellular flux analyzer product details the xf96 extracellular flux analyzer is a fullyintegrated instrument that simultaneously measures the two major energy yielding pathways aerobic respiration and glycolysis in a convenient, microplate format. Add 200 l of seahorse bioscience xf96 calibrant ph 7. Extracellular flux an overview sciencedirect topics. Seahorse xfe24 extracellular flux analyzerbased analysis of. For analyzing hpscs, an appropriate study protocol is to set the time to mix for 2 min, wait for 2 min and measure for 4 min. Seahorse bioscience xfe instrument overview and intended use the seahorse bioscience xfe analyzer measures the rate of change of dissolved oxygen and ph in the media immediately surrounding living cells cultured in.
The xfp extracellular flux analyzer is built on innovative and proven xf technology for live cellbased assays of energy metabolism. The xf96 extracellular flux analyzer was developed to provide higher throughput analysis for customers desiring to study mitochondrial function and dysfunction, obesity, diabetes, cancer, ageing disorders, cardiac disease and drug toxicity. Briefly, cells were plated into xf24 v7 or xf96 v3 polystyrene cell culture plates seahorse bioscience, north billerica. High throughput screening of cellular bioenergetics integra. We have 1 seahorse xf96 manual available for free pdf download. Also notable is that the seahorse xf96 extracellular flux analyzer was cited as a 2009 top ten. Live metabolic profile analysis of zebrafish embryos using. The seahorse xfp analyzer is a compact and easytouse bench top instrument for analyzing energy metabolism in live cells. The seahorse extracellular flux xfe analysers provide a convenient.
Seahorse bioscience announces the xf96 extracellular flux. Scanning seahorse bioscience xf96 cell culture microplates. Ocr and ecar rates are key indicators of mitochondrial respiration and glycolysis and these measurements provide a systemslevel view of cellular metabolic function in cultured cells and exvivo samples. Xfp extracellular flux analyzer from agilent cell analysis. The measurement of cellular bioenergetics on live cells enables timeresolved analysis and testing of multiple conditions per assay well. Seahorse xf cell energy phenotype test kit receives top 10 innovation award from the scientist. Cardamonin inhibits breast cancer growth by repressing hif. Released in late 2006, the xf24 analyzer was and continues to be entirely unique to the biomedical research field. Xfe extracellular flux analyzers simultaneously interrogate the two major energy producing pathways of the cell mitochondrial respiration and glycolysis in a microplate, in realtime. Finally, ocr, glycoper and atp production rates of mitochondrial oxidative phosphorylation and glycolysis were determined and analyzed on the agilents seahorse bioscience xf96 extracellular flux analyzer agilent technologies according to the manufacturers instructions and protocols seahorse bioscience, north billerica, ma, usa. Seahorse xf analyzers measure oxygen consumption rate ocr and extracellular acidification rate ecar of live cells in a multiwell plate, interrogating key cellular functions such as mitochondrial respiration and glycolysis. Seahorse bioscience xf96 extracellular flux analyzer w xf. Statistical analyses were performed using sas software, version 9.