Rm2165 rotary microtome

Manufactured by Leica

Sourced in Germany

The RM2165 rotary microtome is a precision instrument designed for the sectioning of biological samples. It features a motorized cutting mechanism that allows for the precise and consistent cutting of thin sections, enabling microscopic analysis of tissue samples.

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Lab products found in correlation

Fluorescence microscope, by Leica (1 mentions) Copper slit grids, by Electron Microscopy Sciences (1 mentions) Microtome, by Leica (1 mentions) Hard plus resin 812, by Electron Microscopy Sciences (1 mentions) Lsm 5 pascal confocal microscope, by Zeiss (1 mentions) Mz16f microscope, by Leica (1 mentions) Eclipse e800, by Nikon (1 mentions)

Close spelling variants of this product

Microtome (1520 citations) Rotary microtome (327 citations) RM2165 (92 citations) RM2165 microtome (33 citations)

7 protocols using rm2165 rotary microtome

Biotin-SH Barrier Assay for Human Hair Follicles

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For TJ barrier investigation of human HFs, biopsies of 0.5 cm² were taken from the donor scalp samples and prepared as previously described by^{1 (link),27 (link)}.
To study barrier functionality, biopsies were labelled with Biotin-SH. Briefly, 50 μl of 2 mg/mL Biotin-SH (557 Da) in PBS containing 1.0 mM CaCl₂ were injected into the dermis and incubated for 2 h at 37 °C. After incubation, biopsies were fixed for 24 h in Formafix®4%, embedded in paraffin, and 5 μm longitudinal sections were cut using a Leica RM2165 Rotary Microtome (Leica Microsystems GmbH, Wetzlar, Germany). Paraffin sections were deparaffinated, rehydrated, and Biotin-SH was visualised by immunohistochemical staining (see below). For TJ opening experiments, HFs were freshly plucked from donor scalps and incubated for 30 min in PBS with and without EDTA (8 mM) prior to the Biotin-SH barrier assay.

Zorn-Kruppa M., Vidal-y-Sy S., Houdek P., Wladykowski E., Grzybowski S., Gruber R., Gorzelanny C., Harcup J., Schneider S.W., Majumdar A, & Brandner J.M. (2018). Tight Junction barriers in human hair follicles – role of claudin-1. Scientific Reports, 8, 12800.

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Bone Histomorphometry in Mice

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For dynamic bone histomorphometric analysis, 7-week-old mice were injected intraperitoneally with calcein green (10 mg/kg) and alizarin red (20 mg/kg) 6 and 2 days before sacrifice, respectively, as described in a previous study^{30 (link)}. The tibiae were isolated, fixed in 4% paraformaldehyde, and embedded in methyl methacrylate. The embedded tibiae tissues were cut into 6-μm-thick sections using a Leica RM2165 rotary microtome with a tungsten blade (Leica Microsystems, Germany) and then imaged under a fluorescence microscope (Leica Microsystems, Germany). For von Kossa staining analysis, the lumbar vertebrae were fixed and embedded in methyl methacrylate. After that, 6-µm-thick vertebrae sections were stained with von Kossa reagent. To evaluate osteoclast parameters, the tibiae were harvested and fixed in 4% paraformaldehyde, followed by decalcification in 10% EDTA for 4 weeks at 4 °C. The specimens were dehydrated and embedded in paraffin. The paraffin sections were then stained for tartrate-resistant acid phosphatase (TRAP) to identify osteoclasts. Bone histomorphometric analyses were conducted using the Bioquant OSTEO II program (Bio-Quant, Inc., Nashville, TN, USA).

Kim H.J., Lee D.K., Jin X., Che X., Ryu S.H, & Choi J.Y. (2022). Phospholipase D2 controls bone homeostasis by modulating M-CSF-dependent osteoclastic cell migration and microtubule stability. Experimental & Molecular Medicine, 54(8), 1146-1155.

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Histological Analysis of Pioneer Roots and Woody Tissues

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Samples of about 3 mm of pioneer roots and 3 × 2 mm of wood from basal stem and taproot wood from seedlings were fixed, dehydrated and embedded in LR White (London Resin Co., Woking, Surrey, UK) according to [44 ]. Semi-thin transverse sections (1 μm thickness) were cut with a Leica RM2165 Rotary Microtome (Leica Instruments, Heidelberg, Germany) and stained with toluidine blue 0 (CI 52040, Merck, Darmstad, Germany) according to O’Brien et al. (1964). Representative sections of two tissue samples per plant and organ part (basal stem and taproot wood) from six independent plants of each seedling were examined and photographed with an Olympus BX-51 microscope (Olympus Imaging Corp., Tokyo, Japan), with a digital camera DP-12 and the Analysis program (Soft Imaging System GmbH, Munster, Germany). The anatomical data correspond to the mean of six independent plants on each rootstock. Three samples per tissue and plant were studied, and the average values were considered as representative of each individual plant. For each sample, values are the mean of three visual fields from three sections.

Rodríguez-Gamir J., Primo-Millo E, & Forner-Giner M.Á. (2016). An Integrated View of Whole-Tree Hydraulic Architecture. Does Stomatal or Hydraulic Conductance Determine Whole Tree Transpiration?. PLoS ONE, 11(5), e0155246.

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Ultrastructural Analysis of Zebrafish Pronephros

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Larval zebrafish injected with MOs and MO-CTRL were fixed at 120 hpf in 1.5% glutaraldehyde/1% paraformaldehyde, 70 mmol/L NaH₂PO₄, and 3% sucrose (pH 7.2). Further preparation of the zebrafish embryos for TEM was performed as stated in previously published protocols³¹. Briefly, the larvae were washed three times in 0.2 mol/L cacodylate buffer and then post fixed in 1% osmium tetroxide for 1 h at room temperature. The specimens were rinsed with cacodylate buffer, dehydrated in a graded ethanol series, infiltrated and then embedded with Epon (Hard Plus Resin 812; Electron Microscopy Sciences, Hatfield, PA) according to the manufacturer’s protocol. Thin-sections of 0.5 and 1 mm were acquired with a Leica RM2165 rotary microtome followed by staining with 0.5% toluidine blue in a 1% sodium tetraborate solution. Identification of the pronephros was performed after toluidine blue staining, semithin (300 nm) and ultrathin (90 nm) sectioning was performed with a Leica Microtome (Leica Microsystems Inc., Buffalo Grove, IL) and transferred onto copper slit grids (Electron Microscopy Sciences, Hatfield,PA). Grids were stained with uranyl acetate (2%) for 30 minutes and lead citrate for 15 minutes with 3 washing steps in between followed by photograph acquisition and evaluation.

Schenk H., Müller-Deile J., Schroder P., Bolaños-Palmieri P., Beverly-Staggs L., White R., Bräsen J.H., Haller H, & Schiffer M. (2019). Characterizing renal involvement in Hermansky-Pudlak Syndrome in a zebrafish model. Scientific Reports, 9, 17718.

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Ultrastructural Analysis of Zebrafish Pronephros

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Larval zebrafish injected with capped mRNA were sampled at 120 h after fertilization and fixed in 1.5% glutaraldehyde/1% paraformaldehyde, 70 mmol/L NaH₂PO₄, and 3% sucrose (pH 7.2). The embryos were washed three times in 0.2 mol/L cacodylate buffer and then postfixed in 1% osmium tetroxide for 1 h at room temperature. The specimens were rinsed with cacodylate buffer, dehydrated in a graded ethanol series, and infiltrated and embedded with Epon (Hard Plus Resin 812; Electron Microscopy Sciences, Hatfield, PA) according to the manufacturer’s protocol. Thin-sections of 0.5 and 1 µm were generated with a Leica RM2165 rotary microtome and stained with 0.5% toluidine blue in a 1% sodium tetraborate solution. When the pronephros was identified on toluidine blue staining, ultrathin (80- to 100-nm-thick) sections of the kidney were cut and mounted on slot grids (Luxel, Friday Harbor, WA). The sections were stained with 2% uranyl acetate in distilled water and contrasted with lead citrate. Sections were viewed and photographed on a JEOL-1230 transmission electron microscope (Eching, Germany) and an attached charge-coupled device camera.

Teng B., Schroder P., Müller-Deile J., Schenk H., Staggs L., Tossidou I., Dikic I., Haller H, & Schiffer M. (2016). CIN85 Deficiency Prevents Nephrin Endocytosis and Proteinuria in Diabetes. Diabetes, 65(12), 3667-3679.

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Spatiotemporal Gene Expression Analysis

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In situ hybridization was done according to the protocol of Thisse and Thisse (2008) (link). Fluorescence in situ hybridization was done as described (Julich et al., 2005 (link)). Antisense probes used were taz, slc20a1a, and pax2a. After in situ hybridization with the taz probe, some embryos were embedded in JB4 (Polysciences, Warrington, PA) and sectioned with a Leica RM 2165 rotary microtome. Whole mount images of taz and slc20a1a in situ hybridization were taken with a Leica MZ16F microscope (Leica, Wetzlar, Germany), and section images were taken with a Nikon ECLIPSE E800 (Nikon, Tokyo, Japan). pax2a fluorescence in situ hybridization images were taken with a Zeiss Pascal LSM5 confocal microscope (Zeiss, Jena, Germany).

Zhang J., Yuan S., Vasilyev A, & Arnaout M.A. (2015). The transcriptional coactivator Taz regulates proximodistal patterning of the pronephric tubule in zebrafish. Mechanisms of development, 138 Pt 3, 328-335.

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Histological Analysis of Gonadal Adipose and Liver

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Gonadal adipose and liver tissues were fixed in 4 % neutral formaldehyde solution. Adipose tissue and liver tissue were subsequently dehydrated in a graded ethanol series (70-100 %) and embedded in paraffin. The tissues were sectioned (4 μm thick) using a Leica RM 2165 rotary microtome (Wetzlar, Germany) and stained with hematoxylin and eosin (H&E). Sections were viewed with an Axioskop 40 (Oberkochen, Germany) and photographed at 100× magnification.

Lee J.H., Lee J.J., Cho W.K., Yim N.H., Kim H.K., Yun B, & Ma J.Y. (2016). KBH-1, an herbal composition, improves hepatic steatosis and leptin resistance in high-fat diet-induced obese rats. BMC Complementary and Alternative Medicine, 16(1), 355.

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