All tissue samples were collected and prepared for histological examination as described previously [7 (link), 24 (link)]. In brief, brains were removed and postfixed in the same fixative for one week and then washed and stored in PBS at 4°C. The brain from each animal was cryoprotected in 30% sucrose in PBS until the tissue had settled to the bottom of the container. Cryoprotected tissues were then embedded in Tissue-Tek (Sakura Finetek USA Inc., Torrance, CA) and serially sectioned in a coronal plane with a Thermo Cryotome (Thermo Fisher Scientific, Inc., Waltham, MA) at 18–20 μm. Every tenth section from each brain was mounted onto a gelatin precoated slide (total of 20 slides for each brain, with 10–12 sections on each slide).
Thermo cryotome
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Thermo Cryotome is a laboratory instrument used for preparing thin sections of frozen biological samples for microscopic analysis. It enables precise and controlled sectioning of frozen specimens at low temperatures, allowing for the preservation of delicate tissue structures.
Automatically generated - may contain errors
3 protocols using thermo cryotome
1
Histological Preparation of Rodent Brains
2
Apoptosis Detection in Skin Samples
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Human scalp samples and mice dorsal skin were embedded in optimal cutting temperature compound (OCT), and 10 μm thick cryosections were obtained using a Thermocryotome (Thermo, USA) and mounted on coated slides (Dako, Denmark). After washing OCT with phosphate buffered saline (PBS), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) kit (Roche, Germany) was used to detect apoptosis cells according to the manufacturer's instructions. Images were obtained using a fluorescence microscope (Olympus, Japan).
3
Cryotomy Preparation for ToF-SIMS Analysis
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Skin samples removed from Franz cells were placed in a plastic block containing the optimum cutting temperature (OCT) gel (VWR International Ltd., Belgium) which is an inert mounting medium for cryotomy that solidifies upon rapid cooling. Therefore, the plastic block containing skin immersed in OCT was placed in a beaker of isopentane pre-cooled with liquid nitrogen to solidify. After solidification, the OCT blocks were wrapped in aluminum foil, placed in an airtight plastic bags and stored at -80 °C. Cryo-sectioning of skin samples were carried out by placing the OCT block in a cryostat chamber (Thermo Cryotome™, UK) at a temperature of -20 °C. The block was allowed to equilibrate within the cryostat chamber for 30 minutes and then sectioned using a steel blade into vertical cross sections of 20 µm thickness. Following this, the cryo-sections were mounted onto polysine microscope adhesion slides (ThermoFisher Scientific) and freeze dried for 1 hour prior to ToF-SIMS analysis.
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