Optimizing Staining Protocols for Laser Microdissection of Specific Cell Types from the Testis Including Carcinoma In Situ 英文参考文献.docVIP

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Optimizing Staining Protocols for Laser Microdissection of Specific Cell Types from the Testis Including Carcinoma In Situ 英文参考文献.doc

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Optimizing Staining Protocols for Laser Microdissection of Specific Cell Types from the Testis Including Carcinoma In Situ 英文参考文献

OptimizingStainingProtocolsforLaserMicrodissection ofSpecificCellTypesfromtheTestisIncluding CarcinomaInSitu SiBraskSonne1.*,MarleneD.Dalgaard1.,JohnErikNielsen1,ChristinaE.Hoei-Hansen2,EwaRajpert-De Meyts1,LiseMetteGjerdrum3,HenrikLeffers1 1DepartmentofGrowthandReproduction,Rigshospitalet,Copenhagen,Denmark,2DepartmentofPediatrics,Rigshospitalet,Copenhagen,Denmark,3Departmentof Pathology,Rigshospitalet,Copenhagen,Denmark Abstract MicroarrayandRT-PCRbasedmethodsareimportanttoolsforanalysisofgeneexpression;however,intissuescontaining many different cells types, such as the testis, characterization of gene expression in specific cell types can be severely hampered by noise from other cells. The laser microdissection technology allows for enrichment of specific cell types. However,whenthecellsarenotmorphologicallydistinguishable,itisnecessarytouseaspecificstainingmethodforthe targetcells.Inthisstudywehavetesteddifferentfixatives,storageconditionsforfrozensectionsandstainingprotocols, and present two staining protocols for frozen sections, one for fast and specific staining of fetal germ cells, testicular carcinomainsitucells,andothercellswithembryonicstemcell-likepropertiesthatexpressthealkalinephosphatase,and one for specific staining of lipid droplet-containing cells, which is useful for isolation of the androgen-producing Leydig cells.BothprotocolsretainamorphologythatiscompatiblewithlasermicrodissectionandyieldRNAofaqualitysuitable forPCRandmicroarrayanalysis. Citation:SonneSB,DalgaardMD,NielsenJE,Hoei-HansenCE,Rajpert-DeMeytsE,etal.(2009)OptimizingStainingProtocolsforLaserMicrodissectionofSpecific CellTypesfromtheTestisIncludingCarcinomaInSitu.PLoSONE4(5):e5536.doi:10.1371/journal.pone.0005536 Editor:PatrickCallaerts,KatholiekeUniversiteitLeuven,Belgium ReceivedJanuary12,2009;AcceptedApril2,2009;PublishedMay14,2009 Copyright: ? 2009 Sonne et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits