A TMA was constructed from the surgically resected primary tumor tissues of patients with READ as previously described (17 (link),19 (link)), as approved by the IRB of CMUH (approval no. CMUH105-REC2-073). Briefly, the tissues were fixed in 10% neutral buffered formalin (Leica Microsystems, Inc.) for 18–24 h at RT, dehydrated with ethanol, infiltrated with paraffin wax, embedded into paraffin at 60°C and then cooled to become formalin-fixed, paraffin-embedded (FFPE) blocks. Next, the sections of FFPE blocks were cut by a microtome (Leica Microsystems, Inc.) onto the slides. The slides were deparaffinization with xylene, stained with hematoxylin for 3 min and eosin for 1 min, and then mounted in resin. Next, the tumor areas were evaluated and marked on hematoxylin and eosin-stained (H&E) slides by a pathologist using light microscopy (Leica Microsystems, Inc.). Finally, the corresponding area was identified, marked, and punched on the matching FFPE block, and then transferred into a paraffin-embedded recipient block for TMA construction using an AutoTiss 10C system (EverBio Technology, Inc.). A single TMA block comprised a maximum of 60 cores, 2 mm in diameter, and the sections were cut by a microtome (Leica biosystems, Inc.) and mounted on capillary-gap slides (Dako, Inc.).
Microtome
Manufactured by Leica
Sourced in Germany, United States, Japan, China, United Kingdom, Italy, Israel, Australia, France, Canada, Switzerland, Austria, Brazil, Sweden
A Microtome is a precision instrument used to cut extremely thin sections of material, typically for microscopic examination. It operates by moving a sample through a sharp blade, producing uniform slices of the desired thickness. The core function of a Microtome is to enable the preparation of high-quality samples for various microscopic techniques.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Merck Group (26 mentions)
Lsm 780 confocal microscope, by Zeiss (22 mentions)
Cryostat, by Leica (21 mentions)
Tissue processor, by Thermo Fisher Scientific (13 mentions)
Application suite, by Leica (13 mentions)
Paraffin, by Leica (10 mentions)
Superfrost plus slide, by Thermo Fisher Scientific (10 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (9 mentions)
Optical microscope, by Olympus (9 mentions)
Eclipse 80i, by Nikon (9 mentions)
Paraplast, by Merck Group (9 mentions)
Tissue processor, by Leica (9 mentions)
Hematoxylin, by Merck Group (9 mentions)
Zoletil 50, by Virbac (8 mentions)
Image pro plus, by Media Cybernetics (8 mentions)
Historesin, by Leica (8 mentions)
Tp1020, by Leica (7 mentions)
Xylene, by Merck Group (7 mentions)
Permount, by Thermo Fisher Scientific (7 mentions)
Calcein, by Merck Group (7 mentions)
1 520 protocols using microtome
1
Tissue Microarray Construction for Rectal Adenocarcinoma
2
Tissue Harvesting and Preservation Protocol
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At a predetermined time point, after anaesthesia, the rat diaphragm was cut, and the pericardium was opened. After blood collection from the left apex, 5 mm pieces of tissue from the hypothalamus, adrenal glands, spleen and T3 spinal cord were quickly collected from rats in each group, placed in cryopreservation tubes, frozen in liquid nitrogen, and stored in a low-temperature freezer at − 80 °C. Other rats were perfused with 150 mL of sterile 0.9% normal saline and 300 mL of 4% paraformaldehyde (Biosharp, Beijing, China) after blood collection, and the tissues mentioned above were placed in paraformaldehyde overnight. After dehydration in xylene and an alcohol gradient and embedding in paraffin, the samples were cut into 5 µm continuous sections with a microtome or incubated in a sucrose gradient (10%, 20%, and 30%), embedded in OCT compound, and cut into 20 µm continuous frozen sections with a microtome (Leica, Germany).
3
Preparing Paraffin-Embedded Tissue Samples
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Eight to 10 μ m thick paraffin-embedded tissue wax curls were cut from paraffin blocks using a microtome (Leica) with a prion-free microtome blade and prion-free forceps for every FPE sample to avoid cross contamination. Paraffin-tissue curls were subjected to a series of xylene and graded alcohol washes (100%, 95%, and 70%) to remove paraffin and rehydrate tissues similar to standard histological methods. Following alcohol treatments rehydrated FPE tissues were washed with 1 × PBS. Rehydrated FPE tissue was homogenized in at 10% weight per volume by manual disruption or a bead homogenizer as described above.
4
Flower Developmental Morphology Analysis
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Flower buds at different developmental stages were collected and fixed in FAA fixative solution at 4 ℃ overnight. The samples were dehydrated through ethanol series (50% –70%–85%–95%–100%). For histological analysis, the flower samples were embedded in Paraplast® (P3558, Sigma-Aldrich, St. Louis, MO, USA) and 10 μm sections were made by a Leica microtome using a Leica microtome (Leica) and were briefly stained by 0.05% toluidine blue. For SEM analysis, the flower samples after dehydration were subjected to critical point drying in liquid nitrogen and coated with gold, then were examined under an electronic microscope (Zeiss Merlin Compact, Oberkochen, Germany).
5
Lung Cancer Area Quantification
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Lungs fixed with 10% neutral‐buffered formalin were embedded in paraffin and cut into 5‐μm sections using a microtome (Leica Biosystems, Wetzlar, Germany). The sections were stained with hematoxylin (FUJIFILM Wako Pure Chemical Corp.) and eosin (Sakura Fineteck Japan Co., Ltd., Tokyo, Japan) (H&E) or May‐Grunwald staining solution (Merck KGaA, Darmstadt, Germany) and Giemsa staining solution (Merck KGaA) and analyzed for the lung area and cancer area using WinROOF2015 (MITANI Corporation, Fukui, Japan). The cancer area in the lung was calculated as the lung cancer area ratio (%) = lung cancer area/lung area × 100.
6
Laser Capture Microdissection of Gastric Tissue
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LCM was performed for fresh untreated ESD specimens as described previously [13 (link)]. Briefly, specimens were embedded in Tissue-Tek OCT medium (Sakura, Tokyo, Japan), ten serial sections per specimen were cut using a microtome (Leica Biosystems, Deer Park, IL, USA), and transferred to PALM membrane 1.0 PEN slides (Zeiss Microimaging, Munich, Germany). Slides were then stained with hematoxylin and eosin, and coated with Liquid Cover Glass N (Carl Zeiss, Germany). Gastric mucosa (GM), IM, DP, and gastric tumor (GT) cells were delineated using PALM Robosoftware (Zeiss Microimaging), cut out, and collected into 0.5-mL adhesive-cap tubes using a PALM LCM System (Zeiss Microimaging). Genomic DNA of the captured cells was isolated using QIAamp DNA Micro Kit (QIAGEN, Valencia, CA, USA) and its concentration was quantified using PicoGreen dsDNA Quantitation Kit (Molecular Probes, Eugene, OR, USA).
7
Whole-mount Immunofluorescence of Cochlea
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Whole-mount immunofluorescence of cochlea was performed as described previously.30 (link) In brief, isolated cochleas were obtained by microdissection. Tissues were fixed by submersion in 4% formaldehyde at 4°C overnight. After washing twice with PBS, the fixed temporal bones were decalcified for 24 h in 10% ethylenediaminetetraacetic acid (EDTA)/PBS. For paraffin sectioning and H&E staining, serial dehydration of the tissues was performed with ethanol and xylene. Then, the tissues were either embedded in paraffin for standard histological examination or dissected into smaller pieces and permeabilized with 0.1% Triton X-100 for whole-mount immunofluorescence. The paraffin blocks were sliced into 5-μm-thick sections in the midmodiolar plane using a microtome (Leica Biosystems, Nußloch, Germany). Deparaffinization was performed on the sections and whole-mounted tissues with a series of washes with xylene, ethanol, and PBS. After incubation in tris-sodium citrate at 95°C for antigen retrieval, the tissues were blocked with 10% donkey serum and incubated with target-specific primary and secondary antibodies at 4°C overnight. The samples were then mounted with mounting solution (Sigma-Aldrich) and viewed under an LSM780 confocal microscope (Zeiss, Jena, Germany).
8
Testicular Histomorphometry and Germ Cell Analysis
9
Histological Analysis of COX-I and COX-2-KO Brains
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For histology, V, COX-I and COX-2-KO animals were anesthetized with 30 mg/kg Zoletil 50 (Virbac, France) with 0.1 M phosphate-buffered saline (PBS; pH 7.4) delivered via transcardial perfusion followed by 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4). The brains were removed and post-fixed in the same fixative for 12 h. The brain tissues were then dehydrated with graded concentrations of alcohol before embedding in paraffin. Serial sections (3-µm thick) were cut using a microtome (Leica Biosystems, Germany) and mounted onto silane-coated slides (Muto Pure Chemicals, Japan).
10
Formalin-Fixed Tissue Analog Embedding
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