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Efficacy of ImageJ in the assessment of apoptosis
HelmyandAbdelAzimDiagnosticPathology2012,7:15
/content/7/1/15
RESEARCH
OpenAccess
EfficacyofImageJintheassessmentofapoptosis
ImanMHelmy*andAdelMAbdelAzim
Abstract
Objective:ToverifytheefficacyofImageJ1.43nindeterminingtheextentofapoptosiswhichisacomplexand
multistepprocess.
StudyDesign:CisplatinindifferentconcentrationswasusedtoinduceapoptosisinculturedHep2cells.Cell
viabilityassayandnuclearimageanalysisofstainedHep2cellswereusedtodiscriminateapoptoticcellsandcells
suspectedtobeundergoingapoptosisfromcontrolcellsbasedonparameterssuchasnucleararea,circularity,
perimeterandnuclearareafactor(NAF),inassociationwithvisualmorphology.
Results:Imageanalysisrevealedaprogressiveandhighlysignificantdecreaseinnuclearareafactordetectedin
apoptoticcellsandincellssuspectedofundergoingapoptosiscomparedtothecontrolcells(P-values0.01).
Someoftheotherstudiedparametersshowedalsothesametrend.ThisdecreasewasassumedtoindicateDNA
loss.Imageanalysisresultscorrelatedpositivelyandsignificantlywiththeresultsobtainedbycellviabilityassay(R
=0.958,P-value=0.042).NAFwasthemostreliableparameterinassessmentofapoptosis.
Conclusion:Nuclearareafactorcanbecalculatedusingpowerfulfreeandopen-sourcesoftware.Consequently,a
quantitativemeasureofapoptosiscanbeobtainedthatislinkedtomorphologicalchanges.ImageJ1.43nmay
thereforeprovideausefultoolfortheassessmentanddiscriminationofapoptoticcells.
Virtualslides:Thevirtualslide(s)forthisarticlecanbefoundhere:
http://www.diagnosticpathology.diagnomx.eu/vs/5929043086367338
Keywords:Apoptosis,Hep2cell,Imageanalysis,NAF(nuclearareafactor)
Introduction
evolution,apoptosisleadstocondensationandfragmen-
Apoptosis is an active process of self-destruction, tation ofcell bodies, producing regions populated with
wherebycellsundergophysiologicalcelldeath.Itoccurs unstructuredsmallerobjects[4].
duringdevelopmentandregulationoftissuehomeosta-
Examining cells by microscopy has long been a pri-
sis or as a result of changes in environmental stimuli marymethodforstudying
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