The samples fixed in FAA solution were first dehydrated gradually in 50%, 70%, 90%, and 100% ethanol solutions and then dehydrated with a xylene and ethanol mixture (1/3, 1/1, 3/1; v/v) for 30 min each. Next, the samples were dehydrated twice with a xylene and chloroform mixture (9/1, v/v) for 30 min. After infiltration and embedding with paraffin, 8 μm sections were obtained using a rotary microtome (Model RM2245, Leica, Bannockburn, IL, U.S.A.). Additionally, after paraffin was removed using xylene, the sections were subsequently rehydrated in 100%, 95%, 80%, 50%, and 30% ethanol followed by distilled water. After finally being air dried, the sections were observed under a light microscope (Olympus CX31RTSF, Tokyo, Japan).
Model rm2245
Manufactured by Leica
Sourced in Germany
The Leica RM2245 is a rotary microtome designed for sectioning paraffin-embedded tissue samples. It features a vertical specimen feed and an ergonomic hand wheel for precise specimen advancement. The RM2245 is a versatile and reliable instrument for routine histological procedures in research and clinical laboratories.
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Lab products found in correlation
5 protocols using model rm2245
1
Tissue Dehydration and Embedding Methodology
2
Histological Analysis of Rat Liver
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Fresh portions of the liver from each rat were cut rapidly, fixed in 10 % formalin. The fixed tissues were dehydrated with 70, 80, 90, 95 and 100 % ethanol, cleared with 2 changes of xylene, and impregnated with 2 changes of molten paraffin wax, using an automatic tissue processor (Sakura, Japan). The specimens were embedded and blocked out using an embedding station (Sakura, Japan) and serial sections of (4-5 µm) thickness were cut using a microtome (ModelRM2245, Leica, Germany). An autostainer (Model 5020, Leica, Germany) was used for hematoxylin and eosin staining of the sections. The mounted specimens were examined for alterations in the hepatic tissues of each rat under study using optical microscope (Olympus Microscope BP73 with Digital Camera, Japan).
3
Histological Analysis of Plant Tissues
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For light microscopy analyses, entire stem apices, entire leaf primordia and the middle third of young and mature leaves were fixed in Karnovsky's solution (Karnovsky 1965 , modified using pH 7.2 phosphate buffer), placed in a vacuum chamber to remove the air in the tissues, dehydrated in an ethanol series and embedded in hydroxyethyl methacrylate Leica Historesin® (Heraeus-Kulzer, Hanau, Germany), following the manufacturer's instructions, and sectioned at 5–7 µm thickness on a rotary microtome (Model RM 2245, Leica Microsystems Nussloch GmbH, Nussloch, Germany). For structural analysis, the sections were stained with toluidine blue 0.05 % in citrate–phosphate buffer, pH 4.5 (Sakai 1973 (link)), and mounted in synthetic resin Entellan® (Merck®, Darmstadt, Germany).
4
Histological Analysis of Muscle Samples
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The samples were fixed in a 10% buffered formaldehyde solution for 24 h and the historesin protocol was performed (Leica Mycrosistems®, Wetzlar, Germany) [38 (link)]. Sections were performed using a semiautomatic microtome (Model RM2245, Leica Microsystems®, Wetzlar, Germany) with a thickness of 5 µm. The slides were stained with Osmium Tetroxide and counterstained with 1% Toluidine Blue in distilled water. The sections were analyzed under an optical microscope.
Muscle samples were reduced to cylindrical fragments preserving the muscle belly, wrapped in surgical talc, immersed in liquid nitrogen, and included with an adhesive (Optimal Critical Temperature Tissue-Tek® (O.C.T., Sakura Finetek, Torrance, CA, USA)). Then, samples were kept in a freezer at −80 °C until the ten micrometer-thick histological sections were obtained in a cryostat (Model CM 1850, Leica Microsystems®, Wetzlar, Germany) at −20 °C, which were stained with hematoxylin and eosin (HE).
Muscle samples were reduced to cylindrical fragments preserving the muscle belly, wrapped in surgical talc, immersed in liquid nitrogen, and included with an adhesive (Optimal Critical Temperature Tissue-Tek® (O.C.T., Sakura Finetek, Torrance, CA, USA)). Then, samples were kept in a freezer at −80 °C until the ten micrometer-thick histological sections were obtained in a cryostat (Model CM 1850, Leica Microsystems®, Wetzlar, Germany) at −20 °C, which were stained with hematoxylin and eosin (HE).
5
Leaf Anatomy and Histology of Broad Bean
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The broad bean (V. faba L.) leaves were collected from the mature leaves on the shoot during vegetative growth. The specimens were taken from the leaf between mid-vein and the leaf margin and then fixed in formalin-acetic acid alcohol (FAA) using 70% ethanol. The specimens were gradually dehydrated in a tert-butyl alcohol (TBA) series [74 ] and embedded in paraffin wax (m.p. 56 °C). Sections were cut on a rotary microtome at a thickness of 8–10 μm (Model RM2245, Leica Microsystems). Paraffin was removed with xylol solvent and slides were stained with safranin FCF methanol and fast green, and then mounted in Canada balsam [74 ]. The selected sections were examined and photographed using a light microscope (Model BX51; Olympus Optical).
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