Sliding microtome

Manufactured by Leica Biosystems

Sourced in Germany, United States

The Sliding Microtome is a laboratory instrument used for cutting thin tissue sections for microscopic examination. It features a sliding knife that moves horizontally across the specimen, allowing for the precise and consistent sectioning of samples.

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Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Freezing stage, by Leica Biosystems (1 mentions) Vs120, by Olympus (1 mentions) Sucrose, by Merck Group (1 mentions) Panoramic midi 2 scanner, by 3DHISTECH (1 mentions) Peristaltic pump, by Cole-Parmer (1 mentions) Axio imager widefield fluorescence microscope, by Zeiss (1 mentions) Cm1950 cryomicrotome, by Leica Biosystems (1 mentions) Tissue tek oct compound, by Sakura Finetek (1 mentions) Goat anti mouse antibody conjugated to alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Image pro plus, by Media Cybernetics (1 mentions) Prosensor hd, by Planmeca (1 mentions) Epifluorescent microscope, by Leica camera (1 mentions) Leitz dmrb, by Leica camera (1 mentions) Superfrost plus, by Thermo Fisher Scientific (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Axio imager 2, by Zeiss (1 mentions)

16 protocols using sliding microtome

In Vivo Viral Transduction of Engineered MOR Variants

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The AAV (BrainVTA Co., Ltd., Wuhan, Hubei, China) that overexpressed MOR^wildtype/MOR^Y7.43A was rAAV2-EF1α-EGFP-WPRE-hGH polyA, AAV-PHP.eB vector containing MOR^wildtype/MOR^Y7.43A insert of mice.
The heads of mice were firmly fixed by the thumb and index finger. A 25 µl tip microsyringe was used to slowly inject AAVs (5 µl) into the right cerebral lateral ventricle (at coordinates −1.0 mm mediolateral, −0.5 mm anteroposterior from Bregma; −2.0 mm dorsal–ventral from the skull) (Craft et al., 2005 (link)).
Twenty-one days following virus injection, mice were sacrificed through transcardial perfusion of deeply anesthetized mice using intraperitoneal-injected 1% sodium pentobarbital. Mice were perfused with 0.9% saline followed by 4% paraformaldehyde in 0.1 M PO₄, after which whole-brain dissections were performed. Brains were postfixed overnight in 4% paraformaldehyde in 0.1 M PO₄ followed by incubation for a minimum of 48 h in 30% sucrose in 0.1 M PO4 (Kofoed et al., 2021 (link)). Fifty-micrometer sections were cut on a sliding microtome with a freezing stage (Leica Biosystems, Buffalo Grove, IL, United States) (Rincon et al., 2018 (link)). Sections were collected in a 24-well plate filled with antifreeze (PBS: glycerol: ethylene glycol = 5:2:3) and stored in a refrigerator at −20°C and were scanned using Olympus VS.120.

Tian X., Zhang J., Wang S., Gao H., Sun Y., Liu X., Fu W., Tan B, & Su R. (2022). Tyrosine 7.43 is important for mu-opioid receptor downstream signaling pathways activated by fentanyl. Frontiers in Pharmacology, 13, 919325.

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Tissue Harvesting and Preservation for Transplantation Analysis

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Six months after transplantation, the mice were transcardially perfused with PBS. The brain tissues containing grafts from the left and right brain hemisphere were dissected, cut into two halves each, snap frozen on liquid nitrogen and stored at -80°C for further biochemical analysis. For immunohistochemistry, the mice were transcardially perfused with PBS followed by perfusion with 50 ml of 4% paraformaldehyde (PFA). The dissected brains were post-fixed in the same fixative overnight at 4°C. After fixation, brains were cryoprotected in 30% sucrose (Sigma- Aldrich) for 48 h at 4°C before being cut into 30 μm thick coronal serial sections on a sliding microtome (Leica Biosystems, Wetzlar Germany). The free-floating brain sections were then stored in antifreeze solution at −20°C until immunohistochemical staining.

Pomeshchik Y., Velasquez E., Gil J., Klementieva O., Gidlöf R., Sydoff M., Bagnoli S., Nacmias B., Sorbi S., Westergren-Thorsson G., Gouras G.K., Rezeli M, & Roybon L. (2023). Proteomic analysis across patient iPSC-based models and human post-mortem hippocampal tissue reveals early cellular dysfunction and progression of Alzheimer’s disease pathogenesis. Acta Neuropathologica Communications, 11, 150.

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Histological Analysis of Upper Lip

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The upper lip and its surrounding tissue were fixed in 4% phosphate-buffered paraformaldehyde. Specimens were decalcified using 10% ethylenediaminetetraacetic acid for 21 days at room temperature. Using standard methods, paraffin blocks were prepared, and a series of 5 μm thick tissue sections were cut using a sliding microtome (Leica Biosystems, Wetzlar, Germany). Then, the frontal sections were prepared, followed by staining with Masson’s trichrome.

Takagi T., Yamamoto M., Sugano A., Kanehira C., Kitamura K., Katayama M., Sakai K., Sato M, & Abe S. (2022). Alteration of Oral and Perioral Soft Tissue in Mice following Incisor Tooth Extraction. International Journal of Molecular Sciences, 23(6), 2987.

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Tissue Preparation and Histological Analysis

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Animals were deeply anesthetized with xylazine/ketamine and then perfused through the heart with 0.9% ice-cold saline, followed by 4% PFA (pH 7.4). Brains were rapidly removed from skulls, post-fixed in 4% PFA for 1 day at 4°C, and cryoprotected overnight in the same solution, but now containing 30% (w/v) sucrose. Frozen brains were sectioned into 20-μm-thick coronal sections using a sliding microtome (Leica Biosystems). Slices were collected in a cryoprotectant solution (0.05 mol/L sodium phosphate buffer, pH 7.4, 30% ethylene glycol, 20% glycerol) and analyzed by standard hematoxylin:eosin (H&E) staining protocol. Slides were scanned with a Panoramic MIDI II scanner (3DHISTECH Ltd., Budapest). The analysis of the images and the quantification of the areas with lesions were performed using the CaseViewer 2.2, 64-bit version program (3DHISTECH).

Ferreira J.R., Sucupira I.D., Carvalho G.M., Paiva F.F., Pimentel-Coelho P.M., Rosado-de-Castro P.H., Mourão P.A, & Fonseca R.J. (2023). A Combination of Ex Vivo and In Vivo Strategies for Evaluating How Much New Oral Anticoagulants Exacerbate Experimental Intracerebral Bleeding. TH Open: Companion Journal to Thrombosis and Haemostasis, 7(3), e195-e205.

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Brain Tissue Preparation for Histological Analysis

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Ninety minutes after testing, the animals were deeply anesthetized with thiopental (40 mg/kg) and morphine sulfate (10 mg/mL) and perfused transcardially with a solution of 4.0% paraformaldehyde in 0.1 M phosphate buffer, using a peristaltic pump (Cole Parmer). Brains were removed, placed in paraformaldehyde for 3 h, and then transferred to a 30% sucrose/0.1 M phosphate buffer at 4 °C. Frozen whole brain coronal sections (40 mm thick) were sliced with a sliding microtome (Leica Biosystems).

Antunes G.F., Gouveia F.V., Rezende F.S., Seno M.D., de Carvalho M.C., de Oliveira C.C., dos Santos L.C., de Castro M.C., Kuroki M.A., Teixeira M.J., Otoch J.P., Brandao M.L., Fonoff E.T, & Martinez R.C. (2020). Dopamine modulates individual differences in avoidance behavior: A pharmacological, immunohistochemical, neurochemical and volumetric investigation. Neurobiology of Stress, 12, 100219.

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Paraffin and Frozen Sectioning of Fetal Tissues

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We fixed fetal tissues in 4% phosphate-buffered paraformaldehyde (PFA). When we made the paraffin blocks, we decalcified the specimens using 10% ethylenediaminetetraacetic acid (EDTA) for 7 days at room temperature. We prepared the paraffin blocks using standard methods and cut a series of 5-μm-thick tissue sections using a sliding microtome (Leica Biosystems, Wetzlar, Germany). When we made the frozen blocks, we incubated the tissue samples overnight in 30% sucrose in phosphate-buffered saline (PBS). Then, we embedded the tissue samples in Tissue-Tek OCT compound (Sakura Finetek, Japan). Next, we cut 15-μm-thick sections using a CM1950 cryomicrotome (Leica Biosystems, Wetzlar, Germany) and stained the sections with Hoechst 33342 (1:1000 dilution; Thermo Fisher Scientific, Waltham, MA, USA) to visualize the nuclei using an Axio Imager wide-field fluorescence microscope (Zeiss, Oberkochen, Germany). We analyzed the images using ImageJ (NIH, Bethesda, MD, USA). For 3D reconstruction, we loaded the digital images of the hematoxylin and eosin (H&E)-stained serial sections into Amira (Visage Imaging, Inc., Richmond, Australia) by using a voxel size appropriate for the section thickness.

Nagakura R., Yamamoto M., Jeong J., Hinata N., Katori Y., Chang W.J, & Abe S. (2020). Switching of Sox9 expression during musculoskeletal system development. Scientific Reports, 10, 8425.

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Quantification of Amyloid Plaques in Mice

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Mice were deeply anesthetized with 0.14 g/kg sodium pentobarbital, intracardially perfused with PBS, and fixed using 4% paraformaldehyde. Serial coronal sections (35 μm) were cut with a sliding microtome (Leica Biosystems, Buffalo Grove, IL, USA) and stained as freely floating sections. For examination of amyloid plaques, sections were incubated in 88% formic acid at room temperature for 8 minutes. After a washing step in PBS at pH 7.4, sections were blocked in PBS containing 5% bovine serum albumin and 0.5% Triton X-100 for 2 h at 37 °C, then incubated with mouse anti-Aβ antibody (1:1000, Covance 6E10; BioLegend, San Diego, CA, USA) overnight at 4 °C. After further PBS washing, sections were incubated with goat antimouse antibody conjugated to Alexa Fluor 488 (1:500; Thermo Fisher Scientific, Waltham, MA, USA) for 2 h at 37 °C and mounted on (3-aminopropyl)triethoxysilane-coated glass slides. Z-stack images were taken using a Zeiss Pascal (Carl Zeiss Microscopy, Jena, Germany) or Nikon A1 (Nikon Instruments, Tokyo, Japan) laser scanning microscope with a ×20 lens objective. Results were quantified using Image-Pro Plus (Media Cybernetics, Rockville, MD, USA).

Sang S., Pan X., Chen Z., Zeng F., Pan S., Liu H., Jin L., Fei G., Wang C., Ren S., Jiao F., Bao W., Zhou W., Guan Y., Zhang Y., Shi H., Wang Y., Yu X., Wang Y, & Zhong C. (2018). Thiamine diphosphate reduction strongly correlates with brain glucose hypometabolism in Alzheimer’s disease, whereas amyloid deposition does not. Alzheimer's Research & Therapy, 10, 26.

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Carious Tooth Specimen Preparation

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After extraction, the teeth were immediately fixed in 4% neutral buffered formalin (VWR) for a minimum of 24 h and prepared for macroscopic photography and histological processing according to the previously published protocol (Demant et al., 2021 (link)). Briefly, a radiograph was first obtained of each specimen in a buccal‐lingual direction using a Planmeca Prosensor. HD, Planmeca ProX intraoral X‐ray unit and the Planmeca Romexis software (Plandent). The exposure time was 0.6 s, with 6KV and 8 mA as parameters. Each tooth was then sectioned in half through the central part of the occlusal cavitated lesion in a mesio‐distal direction, using a high‐precision saw (MIKRO‐TRENN). Each sectioned half was photographed, and the specimens were subsequently decalcified in 10% EDTA (pH 6.9) and processed for histology. Each macroscopically sectioned half was sectioned serially into 4 μm sections, starting from the central part of the carious lesion until complete exhaustion of the specimens. The number of obtained sections varied greatly from specimen to specimen due to differences in tooth type and section plane. The sectioning procedure was carried out using a sliding microtome (Leica Biosystems). A detailed description of the tissue processing methods has been described elsewhere (Demant et al., 2021 (link)).

Demant S., Schoenmaker T., van Erck S.M., Dabelsteen S., de Vries T.J, & Bjørndal L. (2022). Intra‐pulpal connective tissue formation and the advanced carious lesion: Is chondrogenesis and heterotopic ossification a response to pulpal inflammation?. International Endodontic Journal, 55(11), 1212-1224.

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Neuroanatomical Mapping of ChR2 Expression

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After behavioural testing, mice were deeply anesthetized with pentobarbital (Fatal Plus, 390 mg/ml) and transcardially perfused with 1x phosphate buffered saline (PBS) followed by 4% paraformaldehyde. Brains were removed, placed in vials containing 4% paraformaldehyde overnight at 4 °C, and were then transferred to a 20% (1 day) and then 30% sucrose solution until brains had sunk and ready for slicing. A sliding microtome (Leica Biosystems) was used to cut 30 µm coronal sections, which were subsequently slide mounted and imaged with an epifluorescent microscope (Leica) to identify location of ChR2 virus and optical fiber placement. Only data from mice with correct virus and fiber placement were used for analysis.

Lind E.B., Sweis B.M., Asp A.J., Esguerra M., Silvis K.A., David Redish A, & Thomas M.J. (2023). A quadruple dissociation of reward-related behaviour in mice across excitatory inputs to the nucleus accumbens shell. Communications Biology, 6, 119.

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Histological Examination of Liver and Adipose Tissue

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Liver and retroabdominal adipose tissue samples were taken from each mouse, fixed with a 10% neutral buffered formalin solution, and the samples were embedded in paraffin blocks and sectioned at 5 μm thickness with sliding microtome (Leica Biosystems). The specimens were stained with hematoxylin–eosin (H&E) staining. In brief, the specimens were deparaffinized using xylene and hydrated by gradually decreasing the concentration of ethanol. The specimens were then stained with Mayer's hematoxylin solution for 4 min, followed by color development in running water. Furthermore, they were stained with Eosin solution for 4 min and dehydrated by gradually increasing the concentration of ethanol. After clearing the specimens with xylene, they were cover slipped using a mounting medium.

Nakano T., Sasaki Y., Norikura T., Hosokawa Y., Kasano M., Matsui‐Yuasa I., Huang X., Kobayashi Y, & Kojima‐Yuasa A. (2023). The suppression of the differentiation of adipocytes with Mallotus furetianus is regulated through the posttranslational modifications of C/EBPβ. Food Science & Nutrition, 11(10), 6151-6163.

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