Vibratome Sectioning of Whole Murine Kidney: A Technique to Generate Thin Sections of Murine Kidney
Trascrizione
Use blunt-ended forceps to gently grasp and remove the prepared kidney from ice-cold KRB solution. Immediately place the kidney on absorbent paper for approximately 2 to 4 seconds to remove excess external moisture. Gently roll the kidney across the absorbent paper to ensure that all sides of the parenchyma have dried so that there is optimal adhesion of the kidney to the vibratome stage.
Immediately apply a thin layer of cyanoacrylate glue to the base of the vibratome specimen plate and use blunt-ended forceps to place the kidney, ureter side down, on the area covered in glue. Gently apply downward pressure to the top of the kidney with the flat edge of the forceps for approximately 10 to 20 seconds to dry the glue.
Firmly secure the specimen plate to the bottom of the buffer tray and adjust the level of KRB solution so that the top of the kidney is fully immersed. For automatic vibratome sectioning, select the start and end positions of the vibratome blade cutting cycle, 0.5 to 1 centimeter clear of the kidney, to ensure that the entire kidney plane is getting sectioned.
Start the automatic cutting process, making sure that the blade makes contact with the kidney. Using forceps, collect sections that are liberated from the kidney and immediately transfer them to individual wells.
