The differential diagnosis of small round cell tumors (SRCT) crucially relies on the synoptic evaluation of morphology, immunohistochemical patterns, and molecular features. Though the implementation of broad RNA sequencing in diagnostic molecular pathology routines has substantially changed the standards of molecular affirmation of diagnoses, fluorescence in situ hybridization (FISH) on formalin-fixed, paraffin-embedded (FFPE) tissue sections is still an elementary tool to provide a rapid molecular corroboration of diagnoses, essentially required for therapeutic decisions. We discuss here the major FISH approaches currently employed in diagnostic molecular pathology, addressing classic Ewing sarcoma and differential diagnoses among SRCT which cannot sufficiently be ruled out by immunohistochemistry. This chapter will approach technical issues but particularly strategies and pitfalls in the interpretation of FISH patterns.
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