The hippocampi of each group were fixed in 2.5% ice-cold glutaraldehyde in 0.1 M PBS at pH 7.4 and osmium tetroxide. After gradient dehydration with ethyl alcohol and acetone, the hippocampi were embedded in epoxy resin Epon 812. Tissues were then cut into ultrathin sections (50 nm) by a CM1900 microtome (Leica, Germany) and stained with uranyl acetate and lead citrate. Samples were viewed in a JEOL JEM 1010 TEM at 80 kV and captured through an AMT XR-16 mid-mount 16 mega-pixel digital camera (Hu et al. 2011 (link)).
Cm1900 microtome
Manufactured by Leica
Sourced in Germany
The Leica CM1900 is a rotary microtome designed for high-quality tissue sectioning. It features a manually operated handwheel for precise specimen advancement and a robust, durable construction.
Automatically generated - may contain errors
Lab products found in correlation
Tissue tek oct compound, by Sakura Finetek (2 mentions)
Axioskop 2 fs plus, by Zeiss (1 mentions)
Rompun, by Bayer (1 mentions)
Ov100, by Olympus (1 mentions)
Quanta 200, by Thermo Fisher Scientific (1 mentions)
Methyl green, by Merck Group (1 mentions)
Ab8448, by Abcam (1 mentions)
Hematoxylin, by Leica (1 mentions)
Nanozoomer digital pathology system, by Hamamatsu Photonics (1 mentions)
Von kossa, by Abcam (1 mentions)
Alizarin red s, by Merck Group (1 mentions)
5 protocols using cm1900 microtome
1
Hippocampal Ultrastructural Analysis
2
Lymphatic Tracking of Nanoparticles
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The mice were anesthetized intraperitoneally using a solution containing 8 mg/mL ketamine (Ketalar®, Panpharma, Fougères, France) and 0.8 mg/mL xylazine (Rompun®, Bayer Pharma, Puteaux, France) at 0.015 mL/g of body weight. The lymph node was examined after intradermal injection into the left forepaw pad with the DTX/FPR-675 Pluronic NPs in an aqueous dispersion (10 μL, 1 mg/mL of DTX) using an NIRF image system. Images for lymphatic tracking were obtained at predetermined time points after NP injection using a 580 nm filtered charge coupled device camera under illumination at 460–490 nm, which was implemented in a high sensitivity imaging system (Olympus OV100) for data acquisition and analysis. In order to evaluate the distribution of NPs in lymphatic tissue, the lymph node (brachial) was obtained from normal mice and tumor-bearing mice and the obtained tissue was embedded in a Tissue Tek® OCT compound (Sakura Finetek, Torrance, CA, USA) and cryosectioned at 8 μm (CM1900 microtome, Leica). The fluorescent images were observed under a fluorescence microscope (Olympus BX51 and Zeiss Axioskop2 FS Plus).
3
Observing Byssus Ultrastructure via SEM
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To observe the microscale ultrastructure of the byssus, SEM was performed. The fresh byssus was embedded in PEG-2000 (Sigma), and 10 μm sections were cut on a Leica CM1900 microtome at −20 °C from the distal to proximal region. The sections were washed thoroughly to remove any remaining PEG and were subsequently transferred to a quartz slide. For SEM (FEI Quanta 200, 15 keV), the samples were first vacuum freeze-dried at −40 °C (Christ alpha 1–2) and were then sputter-coated with a nanoroughened film of gold for 60 seconds. For light microscopy, the aforementioned cut samples were viewed on an Olympus BX 60 equipped with a camera.
4
Histological Characterization of Scaffold Samples
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Post micro-CT analysis, three samples of each scaffold type were rinsed in PBS and then embedded in Tissue-Tek® O.C.T. compound (Sakura Finetek, The Netherlands) and stored at −80°C. Slices measuring 14 µm in thickness were sectioned using a CM1900 microtome (Leica) and mounted to glass slides. Slides were then stained with Hematoxylin (Leica) and Eosin (Thermo Fisher Scientific), Alizarin Red S (Sigma-Aldrich) or Von Kossa (Abcam, Cambridge, UK). Immunohistochemistry was performed using a polyclonal antibody to osteopontin (OPN, ab8448, Abcam) and methyl green (Sigma-Aldrich) with a goat pAB to Rb IgG (HRP) (ab6721, Abcam) secondary antibody in normal goat serum (Jackson ImmunoResearch, West Grove, PA, USA). methyl green (Sigma- Aldrich) was used as a counterstain. All histologically stained slides were analyzed qualitatively and imaged using a NanoZoomer Digital Pathology System (Hamamatsu, Japan).
5
Hippocampal Ultrastructural Analysis
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The hippocampi of each group were xed in 2.5% ice-cold glutaraldehyde in 0.1 M PBS at pH 7.4 and osmium tetroxide. After gradient dehydration with ethyl alcohol and acetone, the hippocampi were embedded in epoxy resin Epon 812. Tissues were then cut into ultrathin sections (50 nm) by a CM1900 microtome (Leica, Germany) and stained with uranyl acetate and lead citrate. Samples were viewed in a JEOL JEM 1010 TEM at 80 kV and captured through an AMT XR-16 mid-mount 16 mega-pixel digital camera (Hu et al,2011 ).
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