Microtome instrument

Manufactured by Leica

Sourced in Germany

The Leica Microtome is a precision instrument designed for the accurate sectioning of materials into thin slices or sections. It enables the preparation of high-quality samples for microscopic examination across various scientific and research disciplines.

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Lab products found in correlation

Lsm 710, by Zeiss (2 mentions) Vision lite 2.0 ml, by Optika (1 mentions) Positively charged microscope slides, by Avantor (1 mentions) Cy3 fluorochrome, by Jackson ImmunoResearch (1 mentions) Vectashield mounting medium with dapi, by Vector Laboratories (1 mentions) Bx51 light microscope, by Olympus (1 mentions) Dm750, by Leica (1 mentions) Dfc295, by Leica (1 mentions)

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Microtome (1520 citations)

6 protocols using microtome instrument

Tracing Nanoparticle Biodistribution in Zebrafish

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Zebrafish were anesthetized with 2-phenoxyethanol and injected with 2 µl contain 5 µg/g nanoparticles of FITC conjugated G-AuNPs via intraperitoneal injection. After 4 h, 24 h and 48 h of injection, fishes were sacrificed and digestive system was collected. Organ was fixed with 4% paraformaldehyde and 4% glutaraldehyde followed by dehydration with the gradient increase of ethanol (75%, 95%, 100%) for 30 min each, further with xylene for 1 h and then fixed in paraplast. Blocks were stored in −10 °C for 12 h before proceeding to section. 10 µm thicknesses of sections were cut by using Leica microtome instrument and sections were collected on PLL-coated glass plates. The sections were washed with xylene to remove excess of paraplast. After drying, sections were fixed with mounting media. Sequestration of AuNPs in digestive system was analyzed by confocal fluorescence microscopy using CLSM (Zeiss LSM 710) microscope. The excitation wavelength was 450 nm, detection wavelength was 510 nm and 25 X objective was used to image digestive system sections.

Sangabathuni S., Murthy R.V., Chaudhary P.M., Subramani B., Toraskar S, & Kikkeri R. (2017). Mapping the Glyco-Gold Nanoparticles of Different Shapes Toxicity, Biodistribution and Sequestration in Adult Zebrafish. Scientific Reports, 7, 4239.

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Histopathological Analysis of Mosquito and Moth Larvae

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The NH-EDx-, n-HDa-, n-ODa-, treated and control- fourth instars of A. aegypti and S. litura were fixed in Bouin’s solution overnight, flowingly dehydrated, and mounted in wax blocks using paraffin. Larval blocks of tissue were segmented using the microtome instrument (Leica, Germany), and the active sections were mounted on sterile microscopic glass slides, stained with eosin and hematoxylin, and observed for histopathological changes and photographed using light a microscope (Optika vision lite 2.0 ML) pre-connected with a laptop. Midgut cells of A. aegypti and S. litura were photographed, and the changes in the midgut cells of treated larvae were observed and matched with control [6 (link)].

Amala K., Karthi S., Ganesan R., Radhakrishnan N., Srinivasan K., Mostafa A.E., Al-Ghamdi A.A., Alkahtani J., Elshikh M.S., Senthil-Nathan S., Vasantha-Srinivasan P, & Krutmuang P. (2021). Bioefficacy of Epaltes divaricata (L.) n-Hexane Extracts and Their Major Metabolites against the Lepidopteran Pests Spodoptera litura (fab.) and Dengue Mosquito Aedes aegypti (Linn.). Molecules, 26(12), 3695.

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Histopathological Evaluation of Ovarian Samples

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To perform the histopathological examination, ovarian samples were obtained every two weeks after PRP injection for two months. For this purpose, three rats were randomly selected per group. Rats were humanely euthanized (overdose of ketamine and xylazine) and left ovary was removed, rinsed in phosphate-buffered saline (PBS) solution, and fixed in 10% formalin (Merck). Right ovaries were sampled for gene expression evaluations and stored in liquid nitrogen until use.
For histopathological examination, the specimens were embedded in paraffin, and three consecutive sections of 5 μm thickness prepared using a microtome instrument (Leica). Then, the sections were stained with hematoxylin and eosin (H&E) staining solution. Any pathological response and structural changes were monitored in the ovarian sections. The numbers of primary, secondary, and antral follicles were recorded and compared with the control rats.

Ahmadian S., Sheshpari S., Pazhang M., Bedate A.M., Beheshti R., Abbasi M.M., Nouri M., Rahbarghazi R, & Mahdipour M. (2020). Intra-ovarian injection of platelet-rich plasma into ovarian tissue promoted rejuvenation in the rat model of premature ovarian insufficiency and restored ovulation rate via angiogenesis modulation. Reproductive Biology and Endocrinology : RB&E, 18, 78.

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Immunofluorescence Staining of Tumor Vasculature

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Tumors extracted from mice were embedded in paraffin, cut on a microtome instrument (Leica) in 5 μm-thick sections that were then placed on positively charged microscope slides (VWR). Slides were de-paraffinized in xylene and rehydrated in a series of ethanol solutions (100%, 95%, 70%, 50%, water). Sections were permeabilized by incubating with PBS-T (PBS, 1% Triton X-100) for 15 min at RT and rinsed with water. Antigen retrieval was performed by incubating the slides for 13 min at RT with Proteinase K (at 20 μg/mL final concentration). Slides were washed with PBS 3 × 3 min and incubated with blocking buffer (5% (v/v) goat serum, 1% BSA, 0.5% Tween20 in PBS) for 60 min at RT. Slides were then incubated overnight at 4°C with a rat antibody against CD31 (at 1:100 dilution; Abcam) in a humid chamber. Slides were washed with PBS-T 3 × 10 min and with PBS for 5 min and incubated with the anti-rat secondary antibody conjugated with Cy3 fluorochrome (at 1:300 dilution; Jackson ImmunoResearch) for 2 h. After washing with PBS-T 3 × 10 min and PBS for 5 min, slides were mounted using VectaShield mounting medium with DAPI (Vector Laboratories).

Kubala M.H., Punj V., Placencio-Hickok V.R., Fang H., Fernandez G.E., Sposto R, & DeClerck Y.A. (2018). Plasminogen activator inhibitor-1 promotes the recruitment and polarization of macrophages in cancer. Cell reports, 25(8), 2177-2191.e7.

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Histological Assessment of Ovarian Follicles and Fibrosis

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Formalin-fixed ovarian specimens were embedded in paraffin after dehydration in alcohol series and cut to a thickness of 5 μm using a microtome instrument (Leica). Hematoxylin and eosin (H&E) staining was performed to study the numbers and the quality of follicles at different developmental stages and corpus luteum (CL) [40 ]. Masson’s trichrome staining was also executed to evaluate collagen fiber deposition as a sign of tissue fibrosis [41 ]. After staining, follicular populations and the presence of collagen fibers were evaluated under an Olympus BX-51 light microscope and recorded using a digital camera.

Nazdikbin Yamchi N., Alizadeh Ashrafi M.M., Abbasi H., Amjadi F., Geranmayeh M.H., Shirazi R., Tamadon A., Rahbarghazi R, & Mahdipour M. (2022). Classical music restored fertility status in rat model of premature ovarian failure. BMC Complementary Medicine and Therapies, 22, 290.

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Histological Analysis of Tissue Samples

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After obtaining the paraffin blocks with the tissue samples, they were cut to a thickness of 5 µm using the Leica microtome instrument. After 48 h on a slide, histological analysis was performed using hematoxylin-eosin (HE) and Masson’s trichotomy (MT) stains. Histological images were captured using a Leica DFC 295 camera and a Leica DM750 optical microscope. Image interpretation was performed using Lecia Microsystems 4.12.0 software provided by Leica Microsystem (Mannheim, Germany).
Ethical approval and supervision were performed by the Biomedical Experimentation Animal Ethics Committee (CEAS) of the Universidad del Valle. The study followed the “Animal Research: Reporting of In Vivo Experiments” (ARRIVE) guidelines [41 (link)]. No intraoperative or postoperative complications or deaths of biomodels occurred during the research. Control material was not used to avoid additional discomfort to the animals, as the aim was to compare the biological response to the four formulations. The inclusion criteria considered only sex, age, and weight, while discontinuation criteria included any situation affecting the animals’ welfare, such as intraoperative or postoperative complications.

Castro J.I., Araujo-Rodríguez D.G., Valencia-Llano C.H., López Tenorio D., Saavedra M., Zapata P.A, & Grande-Tovar C.D. (2023). Biocompatibility Assessment of Polycaprolactone/Polylactic Acid/Zinc Oxide Nanoparticle Composites under In Vivo Conditions for Biomedical Applications. Pharmaceutics, 15(9), 2196.

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