Liquid cover glass n

LCM was performed for fresh untreated ESD specimens as described previously [13 (link)]. Briefly, specimens were embedded in Tissue-Tek OCT medium (Sakura, Tokyo, Japan), ten serial sections per specimen were cut using a microtome (Leica Biosystems, Deer Park, IL, USA), and transferred to PALM membrane 1.0 PEN slides (Zeiss Microimaging, Munich, Germany). Slides were then stained with hematoxylin and eosin, and coated with Liquid Cover Glass N (Carl Zeiss, Germany). Gastric mucosa (GM), IM, DP, and gastric tumor (GT) cells were delineated using PALM Robosoftware (Zeiss Microimaging), cut out, and collected into 0.5-mL adhesive-cap tubes using a PALM LCM System (Zeiss Microimaging). Genomic DNA of the captured cells was isolated using QIAamp DNA Micro Kit (QIAGEN, Valencia, CA, USA) and its concentration was quantified using PicoGreen dsDNA Quantitation Kit (Molecular Probes, Eugene, OR, USA).

Fresh untreated specimen from the stomach of a patient with early gastric cancer (EGC) was obtained by endoscopic submucosal dissection (ESD) and embedded in Tissue-Tek OCT medium (Sakura, Tokyo, Japan). Using a cryostat (Microtome, Leica, Germany), ten serial sections (8 μm thick) of the frozen specimen were cut onto PALM Membrane Slide 1.0 PEN slides (Zeiss Microimaging, Munich, Germany), stained with H&E and then coated using Liquid Cover Glass N (Carl Zeiss, Germany) for image enhancement and sample protection. The GM, IM, and GT cells were delineated using the PALM Robosoftware (Zeiss Microimaging) and cut into 0.5-mL Adhesive-Cap tubes using a PALM Laser capture microdissection system (Zeiss Microimaging). The genomic DNAs of the captured cells were obtained using a QIAamp DNA Micro Kit (Qiagen, Valencia, CA). The genomic DNA was subjected to electrophoresis on a 0.8% agarose gel; then, the gel was stained with GelRed (Biotium, Fremont, CA), and the DNA concentration was quantified using a PicoGreen dsDNA Quantitation Kit (Molecular Probe, Eugene, OR).