To remove one for inspection is not difficult, but Villeda then had to cut each brain into wafers 1/25th of a millimetre thick using a cryomicrotome, a machine that resembles a benchtop deli slicer.
Tumor slices (20 μm thickness) were obtained using a cryomicrotome (CM 3050S, Leica Microsystems, Wetzlar, Germany) and were placed on Superfrost Plus microscope slides (Menzel-Glaser, Germany).
Sections were cut from mammary glands with visible tumours and from the lung tissue of the MMTV-PyMT animals using a cryomicrotome (CM 3050S, Leica Microsystems, Wetzlar, Germany) and were placed on Superfrost Plus microscope slides (Menzel-Glaser, Braunschweig, Germany).
After the mice were sacrificed, tumors were removed and immediately frozen in dry ice and sectioned using a cryomicrotome (CM 3050S, Leica Microsystems, Wetzlar, Germany) with a thickness of 25 μm.
Twenty µm brain sections including the SCN and 5 µm ocular sections were prepared using a cryomicrotome (CM1900, Leica, Germany).
Serial 20 µm thick coronal sections of frozen tissues at were obtained at −13°C using a cryomicrotome (CM3050S, Leica, Germany).
For in situ TUNEL assays, the fixed tissues were embedded in OCT compound (Sakura Finetech Japan), and retinal cryosections (5 µm thickness) were obtained by using a cryomicrotome.
Human skin sections (16 µm) were obtained using a cryomicrotome at −17°C, mounted on Fisher Superfrost slides, dried overnight in a desiccator at 4°C, and then kept at −80°C until use.
Paraformaldehyde fixed buds were cryosectioned at 30-40 μm thickness using a cryomicrotome Leica CH 1510-1 (Leica Instruments).
8 mm tissue sections were cut using a cryomicrotome (Leica CM3050S) at −19°C.
