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1、ARTICLEdoi:10.1038/nature12107Structuralandmolecularinterrogationofintactbiologicalsystems1,21121,2KwanghunChung,JenelleWallace,Sung-YonKim,SandhiyaKalyanasundaram,AaronS.Andalman,1,211,2111ThomasJ.Davidson,JulieJ.Mirzabekov,KellyA.Zalocusky,JoannaMat
2、tis,AleksandraK.Denisin,SallyPak,11121,2,3,4HannahBernstein,CharuRamakrishnan,LoganGrosenick,VivianaGradinaru&KarlDeisserothObtaininghigh-resolutioninformationfromacomplexsystem,whilemaintainingtheglobalperspectiveneededtounderstandsystemfunction,repr
3、esentsakeychallengeinbiology.Hereweaddressthischallengewithamethod(termedCLARITY)forthetransformationofintacttissueintoananoporoushydrogel-hybridizedform(crosslinkedtoathree-dimensionalnetworkofhydrophilicpolymers)thatisfullyassembledbutopticallytrans
4、parentandmacromolecule-permeable.Usingmousebrains,weshowintact-tissueimagingoflong-rangeprojections,localcircuitwiring,cellularrelationships,subcellularstructures,proteincomplexes,nucleicacidsandneurotransmitters.CLARITYalsoenablesintact-tissueinsituh
5、ybridization,immunohistochemistrywithmultipleroundsofstainingandde-staininginnon-sectionedtissue,andantibodylabellingthroughouttheintactadultmousebrain.Finally,weshowthatCLARITYenablesfinestructuralanalysisofclinicalsamples,includingnon-sectionedhuman
6、tissuefromaneuropsychiatric-diseasesetting,establishingapathforthetransmutationofhumantissueintoastable,intactandaccessibleformsuitableforprobingstructuralandmolecularunderpinningsofphysiologicalfunctionanddisease.Theextractionofdetailedstructuralandm
7、olecularinformationfromanalysiswithoutdisassembly.Butremovinglipidmembranesthatintactbiologicalsystemshaslongbeenafundamentalchallengeprovidestructuralintegrityandretainbiomoleculeswouldinevitablyacrossfieldsofinvestigation,andhasspurredconsiderablete
8、chnolo-damagetissuewithprofoundlossofcellularandmolecularinforma-1–8gicalinnovation.Thestudyofbrainstructure–functionrelation-tion.Thereforetheprovisionofaphysicalframeworkwouldfirstbe9–12shipsinparticularmaybenefitfromintact-systemtools,andin
