By Gloria H. Su
This formidable selection of crucial but novel equipment for pancreatic melanoma examine or melanoma study regularly positive factors a superb solid of authors who're esteemed leaders within the box. The authors supply a wide diversity of tools for molecular, biochemical, pathological, and statistical research of sporadic and familial pancreatic melanoma, equipment that may be utilized not just to simple, but in addition to translational pancreatic examine. subject matters lined contain in vitro mobile cultures, in vivo mouse versions, protein experiences, mutation research, and therapy improvement.
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Extra resources for Pancreatic Cancer: Methods and Protocols (Methods in Molecular Medicine)
4. 5. Isopentane. Dry ice. Ice bucket. Metal bowl, 500-mL or larger volume. Metal or Plexiglas basket corresponding to metal bowl to hold tissue while freezing. 2. Gentle-Jane® Method 1. Gentle-Jane® (Instrumedics) snap-freezing device. 2. Liquid nitrogen. 3. OCT (“optimal cutting temperature”) compound (Tissue-Tek, others), Cryogel (Instrumedics), or other embedding medium. 2. 1. Tissue Disaggregation Methods Solid tissues can be disaggregated to obtain pure cell mixtures ready for further purification and/or live cell experiments or for direct molecular separation and phenotyping.
Viewing the tissue section under the microscope (directly or on a monitor), the investigator activates the infrared laser when the desired ROI is aligned with a targeting laser beam. The infrared laser (980– 1064 nm) pulse causes localized melting of the thermoplastic membrane, and expansion of the molten plastic causes it to contact the tissue adjacent to ROI. The EVA plastic then resolidifies, remaining adherent to the targeted ROI. As a result, areas where the EVA has come into contact with targeted tissue become optically clearer, allowing the user to identify easily tissues that have already been targeted.
6. 7. 8. 9. 10. Cryostat with blade guards. OCT or other embedding compound. Brightfield microscope. Glass slides. 50-mL conical tubes. 16-in. long smooth forceps. Insulated bucket containing liquid nitrogen. Sterile one-sided straight edge blades. Mayer’s hemalum and eosin slide staining setup. Logbook or computerized logging system. 38 Bova et al. 2. Manual Microdissection of Tissue Sections on Glass Slides The method described is a combination of the one in use in the NCI Pathogenetics Unit and the one described by Moskaluk and Kern (40): 1.