Rat right paws were fixed in 10% formalin were then prepared forhistological analysis by hematoxylin and eosin (H&E) staining (Leica, Melbourne, Australia). The fixed tissue was decalcified with 0.5M ethylenediaminetetraacetic acid, pH 7.4 (Sigma-Aldrich, Missouri, USA). The tissue was then embedded in paraffin (Leica, Melbourne, Australia) and cut to a thickness of 4 μm using a rotary microtome (Thermo Fisher, Massachusetts, USA). The slides were then stained with H&E. The histopathological changes of the tissues were then analyzed by light microscopy (Olympus, Tokyo, Japan) at 100x and 400x magnification and images were digitally captured at a resolution of 1360x1024 pixels with an Olympus DP70 digital camera (Olympus, Tokyo, Japan).
Rotary microtome
Manufactured by Thermo Fisher Scientific
Sourced in United States
A rotary microtome is a laboratory instrument used for cutting thin, uniform slices or sections of biological samples, such as tissues or cells, for microscopic examination. It uses a rotating, reciprocating blade to accurately and consistently section the sample material.
Automatically generated - may contain errors
Lab products found in correlation
Oct compound, by Avantor (2 mentions)
Cryostat 3050, by Leica (2 mentions)
Paraffin, by Leica (2 mentions)
Lsm 700 confocal microscope, by Zeiss (2 mentions)
Dp70 digital camera, by Olympus (1 mentions)
Light microscopy, by Olympus (1 mentions)
Ckx41 microscope, by Olympus (1 mentions)
Histostar, by Thermo Fisher Scientific (1 mentions)
Fluorophore conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Excelsior es tissue processor, by Thermo Fisher Scientific (1 mentions)
Histogel, by Thermo Fisher Scientific (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Pannoramic midi slide scanner, by 3DHISTECH (1 mentions)
Avertin, by Merck Group (1 mentions)
Fv3000, by Olympus (1 mentions)
Antifade mounting medium with dapi, by Beyotime (1 mentions)
Superfrost plus microscope slide, by Thermo Fisher Scientific (1 mentions)
Ni u microscope, by Nikon (1 mentions)
Hematoxylin and eosin, by Merck Group (1 mentions)
Neutral buffered formalin, by Thermo Fisher Scientific (1 mentions)
17 protocols using rotary microtome
1
Histological Analysis of Rat Paws
2
Histopathological Assessment of NAFLD
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Tissues fixed with 4% paraformaldehyde were dehydrated gradually with ethanol (70–100%), and then put on paraffin. The tissues were incised at 4 μm thick by a rotary microtome (Thermo, USA) for hematoxylin and eosin (H&E), masson and Sirius red staining. Steatosis, inflammation, ballooning, and fibrosis in liver were assessed with NAFLD activity scoring system. Sirius red staining was carried out to detect collagen, and masson staining was for fibrosis of liver. Monocyte-derived macrophages and lymphocyte were assessed by immunohistochemistry labeled with CD11b antibody and CD45 antibody, respectively. The apoptosis factors BCL-2 and cleaved-caspase 3 were evaluated by using immunohistochemistry with corresponding antibodies.
3
Intestinal Histomorphology in Piglets
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Two piglets per replicate pen were randomly chosen (based on the average weight of each replicate) at the end of the experiment (day 28) and euthanized by electrical stunning followed by exsanguination. Three replicates (6 piglets per treatment, n = 3) were used for the examination of intestinal histology. Two-centimeter-long segments of the duodenum, jejunum, and ileum were taken from the middle of each part and immediately fixed in 10% neutral buffered formalin (Origin Pure Biosci and Tech, Taipei City, Taiwan). All specimens were sectioned at a thickness of 5 μm (3 cross-sections from each sample) on a rotary microtome (Thermo Scientific, Waltham, MA, USA), followed by staining with hematoxylin and eosin. An image analyzer system composed of an Olympus CKX41 microscope (Olympus Corporation, Tokyo, Japan) and the Visiopharm image software (Hørsholm, Denmark) was used for estimating intestinal morphology. The villus length, crypt depth, and villus-to-crypt ratio of each segment were measured randomly on 30 villi in one piglet.
4
Histological Analysis of Alginate Beads
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For histological analysis, alginate beads were fixed in 4% paraformaldehyde for 30 min at room temperature (25°C) and then placed in PBS for 15 min. Fixed beads were dehydrated using a series of increasing ethanol concentrations (50%, 70%, 95%, and 100% [v/v]); 15 min in each ethanol solution. The ethanol was then replaced with xylene by incubating for 15 min three times and subsequently replaced with paraffin-saturated xylene at room temperature overnight. The beads were placed in an oven (60°C) for 20°min, and xylene was completely replaced by liquid paraffin. The samples were embedded and sectioned at 4 µm using a rotary microtome (Thermo Fisher Scientific) and mounted on slides (10149870, Thermo Fisher Scientific). Samples were dried at 37°C overnight prior to staining (Ismail, 2009 ; Siti-Ismail et al., 2012 (link)).
5
Histological processing of human thymic tissue
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Human thymic samples were fixed (for 2 hrs to overnight) in 4% PFA and processed for either cryo- or paraffin-embedding. For cryo-embedding, fixed tissue was equilibrated in sucrose 25% and embedded in O.C.T. compound (VWR). Cryosections (thickness, 7 μm) were cut on a Leica Cryostat 3050. For paraffin-embedding, a Leica PelorisII tissue processor and Sakura Tissue-Tech embedding station were used. Paraffin section (thickness, 3-5 μm) were produced using ThermoFisher rotary microtome.
Cryo- or Paraffin sections were stained with haematoxylin-eosin using an automatic station (Tissue-Tek Prisma) to verify histology of each tissue and subsequently used for immunohistochemistry analysis.
Cryo- or Paraffin sections were stained with haematoxylin-eosin using an automatic station (Tissue-Tek Prisma) to verify histology of each tissue and subsequently used for immunohistochemistry analysis.
6
Histological Techniques for Human Thymic Tissues
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Human thymic tissues and scaffolds samples were fixed (for 2 h to overnight) in 4% PFA and processed for either cryo- or paraffin-embedding. For cryo-embedding, fixed tissue was equilibrated in sucrose 25% and embedded in O.C.T. compound (VWR). Cryosections (thickness, 7 µm) were cut on a Leica Cryostat 3050. For paraffin-embedding, a Leica PelorisII tissue processor and Sakura Tissue-Tech embedding station were used. Paraffin section (thickness, 3–5 µm) were produced using ThermoFisher rotary microtome. Cryo- or Paraffin sections were stained with haematoxylin-eosin using an automatic station (Tissue-Tek Prisma), Masson’s Trichrome Kit (Leica, Raymond A Lamb, BDH Chemicals) and Van Gieson Staining Kit (Millipore-Merck).
7
Histological Processing of Mouse Tissues
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Tissues of the mouse brain and internal organs were fixed with 10% buffered formalin for 14 days and then dehydrated in 70% EtOH prior to automatic processing and paraffinization in an automatic tissue processor (Spin Tissue STP 120, Thermo Scientific) in a series of increasing concentrations of EtOH, three changes of xylene and two changes of liquid paraffin at 60°C. In order to prepare the paraffin blocks, the processed tissue fragments were embedded using an embedding workstation (HistoStar, Thermo Scientific). Paraffin blocks were cut on a rotary microtome (Thermo Scientific): blocks from the brain hemisphere into 8 μm thick slices and from the livers into 5 μm thick slices.
8
Immunohistochemical Staining of Embryoid Bodies
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EBs were washed once in PBS, fixed in 4% PFA overnight at 4°C, and then embedded in prewarmed HistoGel (Thermo Scientific, specimen processing gel). After cooling, the samples were processed and embedded in paraffin in an Excelsior ES tissue processor (Thermo Scientific). The blocks were sectioned at 5 µm on a rotary microtome (Thermo Scientific) and stained with hematoxylin and eosin if necessary. For IF and IHC, unstained slides underwent standard deparraffinization, rehydration, and antigen retrieval in citrate buffer followed by permeabilization in 0.1% Tx100 and blocking in 5% normal donkey serum (IF) or horse serum (IHC). For IF, after primary antibody incubation overnight at 4°C, sections were stained with appropriate fluorophore-conjugated secondary antibodies (Life Technologies), washed again, DAPI-stained, and mounted in ProLong Gold anti-fade reagent (Life Technologies). For IHC, endogenous peroxidases were quenched with H2O2 followed by overnight primary antibody staining and further processing with ImmPRESS and ImmPact DAB kits from Vector Laboratories. IF images were acquired with a Zeiss LSM700 confocal microscope and processed with Zen 2011 SP2 software, while IHC images were acquired and processed with the Pannoramic MIDI slide scanner from 3DHISTECH. Primary antibody information is in Supplemental Table S2.
9
Histological Evaluation of Disc Degeneration
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Disc tissues were fixed in paraformaldehyde (4% w/v) and decalcified in formic acid solution (10% by wt.) for 4 weeks. Then, discs were embedded in paraffin and cut into 4.0 µm sections using a rotary microtome (Thermo Scientific, USA). The slides were stained with hematoxylin and eosin (HE) and Safranin O‐fast green (SO) according to the manufacturer's instructions. Three independent researchers evaluated the degenerative degree according to the Han et al. histological grading scale.[72 (link)
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10
Immunofluorescence Staining of Mouse Brain Sections
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The mice were deeply anesthetized by 2.5% Avertin (Sigma) and perfused with ice-cold 0.1 mol/L phosphate-buffered saline (PBS), followed by 4% formaldehyde in 0.1 mol/L PBS. The cerebrums were removed from the cranial fossae and immersed in 15% and 25% sucrose at 4°C for 24 h in each concentration. Then, those brains were cut at 20 μm on a rotary microtome (Thermo Fisher, Waltham, USA). The sections were permeabilized with 0.25% Triton X-100, blocked with 2% bovine serum albumin, and then incubated with anti-Ki67, anti-NG2, anti-MBP, anti-Olig2, anti-CC1, anti-GFAP, or anti-Iba1 overnight at 4°C. After 3 washes, those sections were incubated with secondary antibodies (Invitrogen, Carlsbad, USA) for 2 h at room temperature. Finally, antifade mounting medium with DAPI (Beyotime, Shanghai, China) was added to each section to decrease the fluorescence quenching. The primary OPCs were fixed in 4% formaldehyde for 15 min and followed a similar protocol. The laser scanning confocal microscope (FV3000, Olympus) was used to obtain fluorescence images. The images were further analyzed by ImageJ v1.8 software (NIH, MD, USA).
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