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定量单分子检测研究进展.docx

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定量单分子检测研究进展 Abstract: Single-moleculedetectionisanimportantresearchfieldinthelifesciencesandhashadanincreasinglysignificantimpactonvariousfieldsinrecentyears.Quantitativesingle-moleculedetectionisparticularlyimportantasitenablestheprecisemeasurementofbiomoleculesinsmallquantities,providingvaluableinformationforbasicresearchandclinicaldiagnosis.Inthisreview,wesummarizerecentprogressinquantitativesingle-moleculedetectionanddiscussthecurrentchallengesandfutureprospectsofthisresearchfield. Introduction: Single-moleculedetection(SMD)hasbecomeanincreasinglyimportanttoolforstudyingbiologicalprocessesinrecentyears.Differentfromtraditionalensemble-basedtechniques,SMDallowsthemeasurementofindividualmolecules,providingvaluableinformationthatcannotbeobtainedusingensemblemeasurements.QuantitativeSMDisparticularlyimportantasitenablesustodeterminetheconcentrationoftargetmoleculeswithunprecedentedprecisionandtomeasuretheirinteractionkineticswithoutthepotentialforinterferencefromothermolecules. ChallengesinquantitativeSMD: DespitethesignificantadvantagesofquantitativeSMD,itisstillchallengingtoachieveaccuratemeasurementsduetoseveralfactors.First,thesmallnumberoftargetmoleculespresentinsamplesmakesthedetectionprocessdifficult.Second,variationsinthefluorescentsignalandsignal-to-noiseratiocanalsoaffecttheaccuracyofthemeasurements.Finally,itischallengingtodevelopsensitiveandspecificprobesthatcanpreciselylabelthetargetmoleculeswithoutaffectingtheirfunction. CurrentprogressinquantitativeSMD: Despitethesechallenges,significantprogresshasbeenmadeinthedevelopmentofSMDtechniquesoverthelastfewdecades.Oneofthemostsignificantadvanceshasbeenthedevelopmentofsingle-moleculefluorescencemicroscopy,whichenablesthevisualizationandquantificationoftargetmoleculesinrealtimewithhighspatialresolution.Severalvariationsofthistechniquehavebeendeveloped,includingtotalinternalreflectionfluorescencemicroscopy(TIRFM),confocalmicroscopy,andsuper-resolutionmicroscopy.Thesetechniqueshavebeenappliedtostudyawiderangeofb