Rm2245 semi automated rotary microtome

Mice were euthanized and perfused with PBS. The quadricep muscle and joint tissues were harvested, transferred into 15 mL Falcon tubes and fixed with 4% PFA on a rotator at 4 °C overnight. The quadriceps muscle tissues were washed with PBS for 1 h the next day and stored in 70% ethanol for paraffin infiltration. The ankle tissues were decalcified in 14% EDTA for 10 days (solution changed every 2 days) after 48 h of PFA fixation and stored in 70% ethanol for paraffin infiltration. Tissues were infiltrated with paraffin using a Leica TP1020 tissue processor and embedded into paraffin blocks using a Myr Tissue Embedding Centre EC 500. The paraffin blocks were sectioned into 5-μm-thick sections using a Leica RM2245 Semi-Automated Rotary Microtome, and sections were transferred onto a water bath set at 37 °C to unfurl any wrinkled sections. The unfurled sections were collected using positively charged slides. The slides were dried overnight before being subjected to deparaffinization and rehydration. The rehydrated slides were then stained with H&E or safranin-O according to the manufacturer’s protocol. The stained slides were imaged with a Leica Aperio AT2 and exported to QuPath analysis software for quantification of cellular infiltrates and analysis of tissue damage.

To compare the absorption and/or evaporation rate of each formula, the absorption/evaporation time of each sample into bovine hoof slices was further evaluated. Before the experiment, bovine hoof tissues were carefully washed and hydrated in distilled water for 24 h. The softened tissues were cut into thin slices using a microtome (RM2245 Semi-Automated Rotary Microtome, Leica, Germany). The thickness of the slices was measured using a micrometer (Mitutuyo, Guipúzcoa, Spain), and slices with similar thickness and no visible faults or fissures were chosen for the study. The thickness of the hoof slices was ranged from 0.20 to 0.25 mm. A drop of sample (approximately 30 μL) was loaded onto a bovine hoof slice using a pipette, and the time required for complete absorption was determined by dry-to-touch in a shaking incubator maintained at 32 °C.

Sixty one clinical specimens (31 NPC tissues and 30 para-tumor tissues) were collected from the Pathology Department of the Second Xiangya Hospital of Central South University (Changsha, China). The NPC tissues were paraffin-embedded with EG1160 Paraffin Embeddly Center (Leica, Germany), and then sliced using RM2245 Semi-Automated Rotary Microtome (Leica, Germany). Paraffin sections conducted to immunohistochemistry and in situ hybridization.

FFPE blocks from CRC and polyps patients were obtained from the Department of Pathology, UKMMC and validated by the pathologist. Only blocks with more than 80% tumor tissues were selected for IHC staining. The blocks were sectioned at 5 μm using Leica RM2245 semiautomated rotary microtome (Leica Biosystems, Germany) and processed using a standard sectioning protocol.

Following harvest, placental tissue samples (GD15, 16 and 21) were fixed in 3 mL 10% neutral buffered formalin and then embedded in paraffin wax. Placental sections (5 μm; Leica RM2245 Semi-automated Rotary Microtome; Leica Biosystems, U.K.) were stained with haematoxylin and eosin (Sigma-Aldrich, U.K.). Whole sections (3 per slide) were imaged using brightfield microscopy (3D-Histech Pannoramic-250 microscope slide-scanner) at the Bioimaging Facility, University of Manchester. Quantification of placental area, comprising the junctional and labyrinth zones, was performed using ImageJ (NIH, U.S.A.) at 1× magnification per section.