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1、CellStainingIntroductionVisualizationofacellwithfluorescentcompoundsprovidesaimportantroleinoxidativemetabolismbygeneratingATPaswidevarietyofinformationfortheanalysisofcellfunctions.anenergysoruce.TheaveragenumberofmitochondriaperVariousactivitiesandstructuresofacellcanbetargetedforcelli
2、sfrom100to2,000.Thoughthetypicalsizeisabout0.5-stainingwithfluorescentcompounds(Fig.16).Themost2mm,theshape,abundance,andlocationofmitochondriacommonlystainedcellcomponentsarecellmembranes,varybycelltype,cellcycle,andcellviability.Therefore,proteins,andnucleotides.Smallneutralmoleculesan
3、dvisualizationofmitochondraisimportant.Sincemitochondriapositivelychargedmoleculescanpassthroughviablecellhaveelectrontransportsystems,theycanbestainedwithmembranesandremaininsideofcells,dependingontheirvariousredoxdyes.MitoRedandRh123readilypassthroughreactivityorhydrophilicity.Negative
4、lychargedmoleculescellmembranesandaccumulateinmitochondria.Thecannotpassthroughviablecellmembranes.PositivelyfluorescenceintensityofRh123reflectstheamountofATPchargedmoleculesareusuallycellmembranepermeablegeneratedinmitochondria.andaccumulateinthemitochondria.Esterisasuitablefunctionalg
5、roupforstainingviablecellsbecauseitcanpassNucleusStainingthroughviablecellmembranes,whereitishydrolyzedbyFluorescentdyeswitharomaticaminoorguanidinegroups,cellularesterasesintoanegativelychargedmoleculeundersuchaspropidiumiodide(PI),ethidiumbromide(EB),physiologicalconditions.Severalfluo
6、resceinanalogswithdiaminophenylindole(DAPI),acridineorange(AO),andestergroupsintheirstructureareavailableforviablecellHoechstdyes,interactwithnucleotidestoemitfluorescence.staining.SuccinimidylestercompoundscanalsobeusedtoEBandPImoleculesintercalateinsidetheDNAdoublehelix.improvethereten
7、tionofthefluorescentderivativewithintheDAPIandHoechstdyemoleculesattachattheminorgroovecell.ThesecompoundsareneutralmoleculesthatpassoftheDNAdoublehelix.Ontheotherhand,AOcanformthroughcellmembranesandcovalentlyconjugatewithcellcomplexeswitheitherdouble-strandedDNAorsingle
