Single-cell mutation analysis of tumors from stained histologic slides.

摘要: Formalin-fixed and paraffin-embedded tissues are a valuable resource for diagnosis research. PCR is one of the most powerful methods retrospective analysis DNA present in fixed tissues. One major problem with molecular tissue samples, however, cellular heterogeneity, ie, large variety cell types usually these specimens can mask cell-specific genetic alterations associated disease. Herein we describe procedure obtaining analyzing single cells recovered from stained histologic sections without risking contamination neighboring cells. An ultraviolet laser microbeam was used to physically destroy surrounding interest. These cells, now freed adjacent were then easily retrieved motorized, computer-controlled micromanipulator molecularly characterized through use PCR-based microanalysis. This accurate microdissection technique, followed by amplification direct sequencing, revealed novel mutation gene coding adhesion molecule E-cadherin tumor that absent epithelial patient early gastric cancer diffuse type. In this form malignancy, lose homophilic cell-to-cell interactions invade connective as mutations have previously been detected advanced diffuse-type carcinoma lines. The study suggests may be an event tumorigenesis. laser-based isolation subsequent characterization individual described herein, allows micrometer-sized precision should prove useful detecting nucleic acid abnormalities underlie cancer, infection,