Intestine samples (about 2 cm segments) were collected from the mid-region of the intestinal segments: jejunum (from the pancreatic loop to Meckel’s diverticulum) and ileum (from Meckel’s diverticulum to the ileocecal junction) of two birds per pen. The samples were washed with 0.1 M phosphate-buffered saline (pH 7.4) and then fixed in Bouin’s solution for 24 h, followed by preservation in 70% (v/v) ethanol for a day. The samples were then progressively dehydrated at increasing concentrations of ethanol, which was then cleared in xylene. The samples were then embedded with paraffin wax and sectioned at 5 µm thickness using Manual Rotary Microtome (Leica Biosystems, Buffalo Grove, IL, USA). Two sections, at different depths, were made for each sample, which was then stained with hematoxylin and eosin. Pictures at ×50 magnification were taken, and the villus height, width, and crypt depth were measured from five random villi from each replicate using Leica software (Leica DM3000, Leica Biosystems, Buffalo Grove, IL, USA).
Manual rotary microtome
Manufactured by Leica Biosystems
Sourced in United States, Germany, United Kingdom
The Manual Rotary Microtome is a laboratory instrument used to cut thin, uniform sections from a specimen for microscopic examination. It employs a rotating mechanism to advance the specimen through a stationary razor-like blade, producing high-quality tissue sections.
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Lab products found in correlation
3 protocols using manual rotary microtome
1
Intestinal Histomorphometry Analysis of Poultry Samples
2
Tibial Bone Sectioning and Vasculature Staining
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Contralateral left tibiae were dissected free of soft tissues. Dehydration was carried out by immersing tissue in a series of ethanol solutions of increasing concentrations until 100% was reached and the tissue could be infiltrated with the embedding material. Bone cutters (Fine Science Tools, California, USA) were used to section just above the tibiofibular junction (region of interest) and to cut the upper region of the tibia off. Following dehydration, bones were soaked for 24 hours in polymethylmethacrylate (PMMA) at 4 °C. PMMA blocks were ground down using a grinder-polisher (MetaServ 250; Buehler, Illinois, USA) so that the cut surface was flat. 10-µm sections were cut out from the proximal end to trim the bone using a manual rotary microtome (Leica Biosystems, Wetzlar, Germany) until the tibiofibular junction was reached. 5-µm sections were cut and one drop of 80% isopropanol solution added to each section to prevent folding. Sections were pressed in an incubator at 37 °C overnight to dry the tissue and stained following either Pentachrome or Giemsa protocols to confirm the presence of blood vessels.
3
Leaf Anatomy and Isotopic Analysis of S. isaloensis
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Herbarium collections were studied at P, K, and TAN herbaria. Tissue harvested from a recently collected herbarium specimen (Razanatsoa et al. 578, K) was used to characterize leaf anatomy and photosynthetic type of S. isaloensis. Leaf pieces of approximately 5 mm in length were rehydrated for 24 hr then fixed in Carnoy's fixative (4:1 EtOH:acetic acid) and embed-ded in Methacrylate embedding resin (Technovit 7100, Heraeus Kulzer GmbH, Wehrhein, Germany). Embedded leaves were sectioned between 6-8 mm thick on a manual rotary microtome (Leica Biosystems, Newcastle, UK) and stained with Toluidine Blue O (Sigma-Aldrich, St. Louis, MO, USA). Stained leaf sections were photographed using microscopy imaging software and a camera mounted on a microscope (CellA; Olympus DP71; BX51, respectively. Olympus, Hamburg, Germany). Images were stitched together (DoubleTake 2.2.9, Echo One, Frederikssund, Denmark) to recreate the continuous width of the whole cross-section.
Dried leaf tissue was prepared for δ 13 C analysis by the University of Sheffield, Faculty of Science biOMICS facility. The δ 13 C value is presented as an isotopic ratio in parts per thousand (per mil, %), reported relative to the isotopic standard Pee Dee Belemnite (PDB).
Dried leaf tissue was prepared for δ 13 C analysis by the University of Sheffield, Faculty of Science biOMICS facility. The δ 13 C value is presented as an isotopic ratio in parts per thousand (per mil, %), reported relative to the isotopic standard Pee Dee Belemnite (PDB).
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