In Situ Hybridization to Somatic Chromosomes in Drosophila

InsituhybridizationwasoriginallydevelopedasatechniqueforvisualizingandphysicallymappingspecificsequencesonDrosophilamelanogasterpolytenechromosomes.Hybridizationtechniquescanalsobeusedtolocalizesequencesonsmaller,diploidchromosomes,suchascondensedmitoticchromosomes.Variationsofthemethodalsoallowthehybridizationofprobestochromosomeswithinintactcellsandtissues,ratherthantochromosomesisolatedfromtheircellularcontextandflattenedonslides.Thisarticlepresentsmethodsforhybridizingfluorescentprobestochromosomesinwhole-mountDrosophilatissues.Thesemethodsallowtheinvestigationofnuclearorganizationevenatstageswherechromosomesaredecondensed(asininterphase)or,forotherreasons,cannotbediscriminatedinthelightmicroscope.Consequently,theyareusefulforaddressingavarietyofcellbiologicalquestions.Inadditiontoenhancingourunderstandingofsomaticchromosomeorganization,thisexperimentalapproachhasalsorevealedinteractionsamongmeioticchromosomesinDrosophilafemales,whichspendmuchofmeiosisinacompactballcalledthekaryosome.Fluorescentinsituhybridization(FISH)methodscanalsobeusedtokaryotypeindividualnucleiusingchromosome-specificMarkers.Withappropriatefixationconditions,hybridizationtochromosomalDNAcanbeperformedinconjunctionwithimmunostaining,allowingthecolocalizationofcellularorchromosomalproteins.

原文：http://cshprotocols.cshlp.org/content/2011/9/pdb.top065540.abstract