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Cellutions 2012V2

Cellutions 2012V2

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The Newsletter for Cell Biology Researchers
The Newsletter for Cell Biology Researchers

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Categories:Types, Research, Science
Published by: EMD Millipore Bioscience on Jul 11, 2012
Copyright:Attribution Non-commercial

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07/11/2012

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Cell
utins
 Vl 2: 2012
The Newsletter rCell Bilgy Researchers
EMD Millire is a divisin  Merck KGaA, Darmstadt, Germany
What killed yur cells?
Multiplexed detection o changing cytokineproles with respect to PBMC stimulation,sepsis and autoimmune diseases
Page 7
LentiBrite™ Lentiviral Biosensors orfuorescent cellular imaging: analysis o thecytoskeleton
Page 11
Cellular characterization o chromatin statevia high throughput ChIP assay
Page 16
REVIEW – Sonic Hedgehog: its dualrole in morphogenic and mitogenicsignaling
Page 21
To subscribe to the quarterly Cellutions newsletter,please visit 
Simlied evaluatin atsis using theMuse™ Cell Analyzer
Page 3
 
2
Product HIGHLIGHt
Nw, exerience it.
 
Smarter cell analysis with yur wnMuse
Cell Analyzer.
Revlutinize the way yu analyze cell viability, atsis andcell cycle with the Muse™ Cell Analyzer. Using miniaturizedfurescence-based detectin, a user-riendly interace andtimized assays, the Muse™ Cell Analyzer rvides werulcell analysis simly, accessibly, and ardably. Exerience thenew Muse™ Cell Analyzer and make smarter, aster and mreaccurate decisins abut yur exeriments.
 
Simly see mre.
San his 2d ba e wih ymbile evie.
 
3
Inin
The degree o apoptosis in a cell population is an importantparameter that contributes to a comprehensive pictureo cell health. Accurate detection and measurement o cellular apoptosis is essential or drug development anddiscovery, or understanding mode o compound action,and or understanding the impact o culture and growthconditions. However, the assessment o cellular apoptosishas been limited due to the requirements or expensiveand complicated instrument platorms, expertise andimproved analytical methods that provide rapid, robust andreproducible apoptosis data. Access to these improvementscan acilitate apoptosis monitoring, thereby enabling theecient, daily execution o cellular research.The Muse™ Cell Analyzer is a novel instrument that enablesmultidimensional cell health analysis on a single platorm.This small, benchtop cell analyzer eortlessly guides usersthrough the acquisition and analysis o samples usinga highly simplied and intuitive touchscreen interacewhich delivers rapid measurements o cell concentration,viability, apoptotic status, and cell cycle distribution
1
. Usingmultiparametric fuorescent detection o individual cellsvia microcapillary fow technology, the system enableshighly sensitive and rapid detection o cellular samplesusing minimal cell numbers. The simplied ormat enablesresearchers o varying backgrounds and experience levelsto obtain a comprehensive picture o cellular health.In this study, we used the Muse™ Cell Analyzer orapoptosis studies using the Muse™ Annexin V & DeadCell Assay, a rapid, no-wash assay or the identicationo apoptotic cells. Jurkat and HeLa cells were treated witha range o cytotoxic and anti-tumor compounds, and theresults rom percentage o live, early, late apoptotic, anddead cells were evaluated. Our results demonstrate thatthe assay provides rapid and sensitive detection o cellularapoptosis and provides quantitative and precise results ona variety o cellular types and compound treatments.
Maeials an Mehs
Assay piniple
Apoptosis, or programmed cell death, is an importantregulator o cell growth and prolieration and ischaracterized by distinct morphological changes. Onekey early aspect o apoptosis is the translocation o phosphotidylserine rom the inner to the outer leafet o the plasma membrane and exposure to outer surace o thecell. This universal phenomenon is independent o species,cell type and induction system and occurs early in theapoptotic process.The Muse™ Annexin V & Dead Cell Assay is based on thedetection o phosphatidylserine (PS) on the surace o apoptotic cells and uses a premixed reagent containingfuorescently labeled Annexin V in combination with adead cell marker, 7-AAD. Annexin V is a Ca
2+
-dependentphospholipid-binding protein that has a high anity orPS, a membrane component normally localized to theinternal ace o the cell membrane. Early in the apoptoticpathway, molecules o PS are translocated to the outersurace o the cell membrane where Annexin V can readilybind to them (Figure 1). Late-stage apoptotic cells showloss o membrane integrity. The membrane-impermeantdye, 7-AAD, is used to distinguish dead cells rom earlyapoptotic cells. The assay can thus distinguish ourpopulations:
•
 Viable cells, not undergoing detectable apoptosis:Annexin V (–) and dead cell marker (–)
•
Early apoptotic and dead cells: Annexin V (+) and deadcell marker (–)
•
Late apoptotic or dead cells: Annexin V (+) and dead cellmarker (+)
•
Cells that have died through non-apoptotic pathway:Annexin V (–) and dead cell marker (+)
Simlied evaluatin  atsisusing the Muse
Cell Analyzer
Asima Khan, Kaheine Gillis, Jlie cl, Kamala tyagaajan
EMD Millire Crratin
Fige 1. 
The cmbineduse  furescent labeledAnnexin V and membrane-imermeant 7-AAD DNA-binding dye can distinguishlive, early attic, and lateattic cells.
Healthy CellApoptotic CellLate Apoptotic Cell

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