Rm2255 rotary microtome

Manufactured by Leica camera

Sourced in Germany

The RM2255 rotary microtome is a precision instrument used for cutting thin sections of biological samples for microscopic examination. It features a sturdy construction, a reliable drive mechanism, and a range of adjustable settings to ensure accurate and consistent sectioning.

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Lab products found in correlation

Tp 1020 sample blocker, by Leica camera (2 mentions) Lsm800 confocal microscope, by Zeiss (2 mentions) Histoclear, by National Diagnostics (1 mentions) Methyl methacrylate, by Fujifilm (1 mentions) Insulin, by Agilent Technologies (1 mentions) Bip c50b12, by Cell Signaling Technology (1 mentions) Bouin s fixative, by Merck Group (1 mentions) R buffer a, by Electron Microscopy Sciences (1 mentions) Anti cdh1, by BD (1 mentions) A1 confocal microscope, by Zeiss (1 mentions) Nanozoomer slider scanner digital pathology system, by Hamamatsu Photonics (1 mentions) 500 mhz nmr spectrometer, by Agilent Technologies (1 mentions) Agilent 1100 hplc system, by Thermo Fisher Scientific (1 mentions) Kta pure system, by GE Healthcare (1 mentions) Hypersil ods c18 column, by Thermo Fisher Scientific (1 mentions) Pearl imaging system, by LI COR (1 mentions) Superdex 200 increase column 10 300 gl, by Wyatt Technology (1 mentions) Smoothened, by Santa Cruz Biotechnology (1 mentions) 3 3 diaminobenzidine (dab), by CWBIO (1 mentions) Arl13b, by Proteintech (1 mentions)

24 protocols using rm2255 rotary microtome

Histochemical Analysis of Plant Stem Sections

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Tissues were fixed for 6 h in an FAA solution (3.7% formaldehyde, 5% acetic acid, 50% ethanol) and subsequently dehydrated through an ethanol series. The tissues were cleared with Histoclear (National Diagnostics) and embedded in paraffin. An RM2255 rotary microtome (Leica) was used to make 8 μm transversal stem sections.
After deparaffinisation, sections were stained with an Alcian Blue 8Gx/Safranin-O solution (0.05% Alcian Blue 8Gx and 0.01% Safranin-O in 0.1 M acetate buffer [pH 5.0]) as described in Østergaard et al. (2006 (link)). Sections were examined under light microscopy.

Stephenson P., Stacey N., Brüser M., Pullen N., Ilyas M., O’Neill C., Wells R, & Østergaard L. (2019). The power of model-to-crop translation illustrated by reducing seed loss from pod shatter in oilseed rape. Plant Reproduction, 32(4), 331-340.

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Nanobody-IL2 Fusion Therapy for Melanoma

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Mice were inoculated on day 0 with 1 M B16F10 cells in 50 μL PBS injected s.c. in the right flank. Mice were treated with 100 μg TA99 (i.p.) on day 6, and PBS or 0.4 nmol nanobody-IL2 fusions (i.t.) on days 6 and 10. On day 12, tumors were excised and fixed in 4% formaldehyde in PBS at RT overnight and paraffin-embedded following standard procedures. Consecutive sections (4 to 6 μm) were prepared using a Leica RM2255 rotary microtome, dried at 60°C for 1 hour. Hematoxylin and eosin (H&E) staining was performed using standard protocols with the help of a Thermo Scientific Shandon Varistain Gemini ES Automated Slide Stainer. A Leica Apeiro AT2 slide scanner was used for image documentation.

Lutz E.A., Jailkhani N., Momin N., Huang Y., Sheen A., Kang B.H., Wittrup K.D, & Hynes R.O. (2022). Intratumoral nanobody–IL-2 fusions that bind the tumor extracellular matrix suppress solid tumor growth in mice. PNAS Nexus, 1(5), pgac244.

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Femur Histomorphometry Analysis Protocol

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After sacrifice, the left femur was isolated, fixed in 70% pure ethanol, and embedded in methyl-methacrylate (Wako Pure Chemical Industries, Osaka, Japan) without bone decalcification. Frontal plane sections (5 μm thick) of the distal metaphysis were cut using a RM2255 rotary microtome (Leica, Wetzlar, Germany) and stained with Villanueva osteochrome bone stain (basic fuchsin, fast green, orange G, and azure II; Merck, Darmstadt, Germany) for bone histomorphometry as previously reported [24 (link)]. Static and dynamic histomorphometric analyses were measured at the Ito Bone Histomorphometry Institute (Niigata, Japan). The following parameters were measured using a semiautomatic graphic system (Histometry RT CAMERA; System Supply Co., Nagano, Japan): BV/TV (%); Tb.Th (μm); Tb.N (N/mm); Tb.Sp (μm); osteoclast surface per bone surface (Oc.S/BS, %); eroded surface per BS (ES/BS, %); number of osteoclasts per BS (N.Oc/BS, N/mm); number osteoblast surface per BS (Ob.S/BS, %); mineralizing surface per BS (MS/BS, %); number of osteoblasts per BS (N.Ob/BS, N/mm); bone formation rate per BS (BFR/BS, μm³/μm²/day), and mineral apposition rate (MAR, μm/day) [30 (link)].

Ju Y.I., Choi H.J, & Sone T. (2022). Effects of Korean red ginseng on three-dimensional trabecular bone microarchitecture and strength in growing rats: Comparison with changes due to jump exercise. PLoS ONE, 17(5), e0267466.

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Histological Analysis of Tumor Samples

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Samples were fixed in 4% buffered formaldehyde for 24 h, washed in PBS, transferred into 70% EtOH in ddH₂O, and then embedded in paraffin and sectioned (5mm thickness) using a Leica RM2255 rotary microtome, by the Brain Tumor Research Center (BTRC) Histology Core, or by the Biobank and Tissue Technology Lab at UCSF. Hematoxylin and eosin staining was performed using standard methods. Stained slides were imaged using an Aperio AT2 slide scanner and data were processed using QuPath software. Total cell counts, Ki67 stains in all tissues, and cleaved caspase-3 stains in RT2 tumors were automated in a blinded fashion; cleaved caspase-3 stains in INS-1 xenografts were quantified manually in a blinded fashion. Antibodies used for immunohistochemistry: BiP [C50B12] (Cell Signaling Technology #3177, 1:200), CD31 (CST #77699 1:100), Chromogranin A [polyclonal] (Cell Marque, 1:4), Cleaved Caspase-3 (CST #9661 1:200), Insulin (DAKO A0564, 1:200), IRE1α (CST #3294, 1:100), Ki67 (Ventana #790–4286, Undiluted), Myc (Sigma M4439, 1:5000), Synaptophysin [LK2H10 clone] (Cell Marque, 1:100).

Moore P.C., Qi J.Y., Thamsen M., Ghosh R., Peng J., Gliedt M.J., Meza-Acevedo R., Warren R.E., Hiniker A., Kim G.E., Maly D.J., Backes B.J., Papa F.R, & Oakes S.A. (2019). Parallel signaling through IRE1α and PERK regulates pancreatic neuroendocrine tumor growth and survival. Cancer research, 79(24), 6190-6203.

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Quantification and Histological Analysis of Lung Metastases

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For quantification of metastatic lesions, mouse lungs were isolated, washed briefly in PBS, fixed in Bouin’s fixative (Sigma) overnight at 4 °C, and stored in 70% ethanol before counting lung nodules. For histological analysis, lungs were fixed in 10% formalin overnight at 4 °C, dehydrated, and embedded in paraffin (Tissue Tek Embedding Station). 3μm sections were cut on a Leica RM2255 rotary microtome. For antigen retrieval, deparaffinized slides were immersed in R-buffer A (Electron Microscopy Services) and boiled in a microwave for 20 min. Samples were blocked and permeabilized for 30 min in blocking buffer (5% normal goat serum, 0.3% TritonX-100 in PBS), followed by primary antibody incubation overnight at 4 °C using anti-CDH1 (1:100, BD Biosciences, 610181). Secondary antibody (1:500, BD Biosciences) was incubated for 1 h at RT. All sections were analyzed using a Nikon A1 confocal microscope and Zeiss fluorescence microscope.

Celià-Terrassa T., Bastian C., Liu D.D., Ell B., Aiello N.M., Wei Y., Zamalloa J., Blanco A.M., Hang X., Kunisky D., Li W., Williams E.D., Rabitz H, & Kang Y. (2018). Hysteresis control of epithelial-mesenchymal transition dynamics conveys a distinct program with enhanced metastatic ability. Nature Communications, 9, 5005.

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Histological Analysis of Gingival Tissue

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Gingival tissues were formalin fixed (4% neutral buffered formalin) for 24 hours at room temperature and further placed in 70% ethanol. Tissues were processed by graded dehydration in alcohols using TissueTek VIP1000 and paraffin embedded. Serial sections of 5μm were obtained using Leica RM2255 Rotary Microtome and tissues were deparaffinized following standard protocol with xylene, and rehydrated with decreasing concentrations of ethanol. Sections of each biopsy were stained with Hematoxylin-Eosin (H&E). The areas of connective tissue under the epithelial basement membrane were analyzed under light microscopy at 200×. Each gingival tissue sample was divided into 10 fields and inflammatory cell infiltrate was determined in each field based on morphological characteristics by a blinded investigator.

Ebersole J.L., Kirakodu S., Novak M.J., Exposto C.R., Stromberg A.J., Shen S., Orraca L., Gonzalez-Martinez J, & Gonzalez O.A. (2015). Effects of aging in the expression of NOD-like receptors and Inflammasome-related genes in oral mucosa. Molecular oral microbiology, 31(1), 18-32.

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Mouse Thigh Tissue Analysis

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The mouse thigh was split into 4 sections between hip and knee, fixed in 10% buffered formalin, and embedded in paraffin. Tissues were cut at a 4 µm thickness using a Leica RM 2255 rotary microtome and the cross-sections were stained with Hematoxylin and Eosin or Prussian blue stain to analyze tissue morphology and locate SPIONs, respectively. Tissue sections were visualized with a NanoZoomer Slider Scanner/Digital Pathology System (Hamamatsu).

Smith C.E., Ernenwein D., Shkumatov A., Clay N., Lee J., Melhem M., Misra S., Zimmerman S.C, & Kong H. (2015). Hydrophilic Packaging of Iron Oxide Nanoclusters for Highly Sensitive Imaging. Biomaterials, 69, 184-190.

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Multimodal Characterization of Polymer-based Formulations

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¹H and ¹³C NMR spectra were recorded on a 500 MHz NMR spectrometer (Varian, Palo Alto, CA, USA). ÄKTA Pure system (GE HealthCare, Chicago, IL, USA) equipped with UV detector and Superdex 200 increase column 10/300 GL, multi-angle light scattering (MALS) detector (Wyatt, Santa Barbara, CA, USA) and differential refractive index (dRI) detector (Wyatt, Santa Barbara, CA, USA) were used to determine the copolymers’ weight average molecular weight (M_w) and number average molecular weight (M_n). Drug content was analyzed on an Agilent 1100 HPLC system (Santa Clara, CA, USA) with a Hypersil™ ODS C₁₈ Column (Thermo Scientific, Waltham, MA, USA). Paraffin-embedded tissue sectioning was done using a Leica RM2255 rotary microtome (Buffalo Grove, IL, USA). Histology slides was scanned by a VENTANA iScanner HT (Tucson, AZ, USA). Immuno-fluorescence labeled slides were analyzed using a ZEISS LSM 800 confocal microscope (Peabody, MA, USA). Live animals were imaged using LI-COR^® Pearl imaging system (Lincoln, NE, USA). The endoscope used for the colitis scoring was purchased from Gradient Lens Corporation^® (Rochester, NY, USA)

Zhao G., Wei X., Wu J., Eichele D.D., Lele S.M., Yang L., Zhang F, & Wang D. (2019). A Macromolecular Janus Kinase (JAK) Inhibitor Prodrug Effectively Ameliorates Dextran Sulfate Sodium-induced Ulcerative Colitis in Mice. Pharmaceutical research, 36(4), 64.

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Tissue and Cell Immunostaining Protocol

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For tissue staining, dorsal skin were fixed with 4% paraformaldehyde, dehydrated in graded ethanol series and embedded in paraffin. Sections of 5 µm thickness were prepared by Leica RM2255 rotary microtome and used for immunostaining after deparaffinization. For cell staining, Ptch^−/− MEF cells cultured on cover-slips were fixed with 4% paraformaldehyde, blocked with 5% BSA for 1 h and incubated overnight with primary antibody. The following antibodies were used for immunostaining: Smoothened (Santa Cruz, sc-166685, 1:100); ARL 13B (Proteintech, 17711-1-AP, 1:100); Gli1 (Proteintech, 66905–1-Ig, 1:200). The secondary antibodies conjugated with Alexa Fluor 488 or 568 were purchased from Invitrogen, those conjugated with horse radish peroxidase (HRP) using DAB as chromogen from Cwbio. Sections were photographed by LSM710 Zeiss confocal microscope and analyzed with Image J software.

Fan J., Li H., Kuang L., Zhao Z., He W., Liu C., Wang Y., Cheng S.Y, & Chen W. (2021). Identification of a potent antagonist of smoothened in hedgehog signaling. Cell & Bioscience, 11, 46.

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Histological Evaluation of Organ Tissues

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Spleen, kidney, and liver tissue samples from the control and treated animals were fixed in 4% formaldehyde and embedded into paraffin. The analysis and evaluation were performed at the Czech Centre for Phenogenomics, Institute of Molecular Genetics of the CAS, Vestec, using Leica ASP6025 automatic vacuum tissue processor for tissue processing, Leica EG1150 H + C embedding station for embedding, and Leica RM2255 rotary microtome for sectioning (2 μm sections). Standard haematoxylin and eosin (HE) descriptive histopathological staining was done using Leica ST5020 + CV5030 stainer and coverslipped.

Chrienova Z., Rysanek D., Oleksak P., Stary D., Bajda M., Reinis M., Mikyskova R., Novotny O., Andrys R., Skarka A., Vasicova P., Novak J., Valis M., Kuca K., Hodny Z, & Nepovimova E. (2022). Discovery of small molecule mechanistic target of rapamycin inhibitors as anti-aging and anti-cancer therapeutics. Frontiers in Aging Neuroscience, 14, 1048260.

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