pubmed-article:16028412 | pubmed:abstractText | Laser microdissection is an essential method for the investigation of the multistep carcinogenic process in the urinary bladder. Reliable detection of tumor-specific alterations which can be compromised by the presence of normal cells, requires microdissection of pure tumor cell populations (>80%) to detect loss of heterozygosity (LOH) by either fluorescence in situ hybridization (FISH) or sequence analysis. Multiple molecular methods need to be performed in the course of studying often-small lesions. This chapter describes in detail the use of laser microdissection, whole-genome amplification by improved primer extension preamplification (I-PEP)-polymerase chain reaction, and subsequent LOH, FISH, and sequencing analysis in the investigation of urothelial tumors and their precursor lesions. The combination of the described methods allows a wide spectrum of molecular investigations of tumor cells and helps to understand the fundamental alterations involved in urothelial carcinogenesis. | lld:pubmed |