Sliding microtome

Manufactured by Thermo Fisher Scientific

Sourced in Germany, United States

The Sliding Microtome is a laboratory equipment used for thinly slicing samples for microscopic examination. It features a sliding mechanism that allows the user to precisely control the thickness of the tissue sections, typically ranging from 1 to 60 micrometers.

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Lab products found in correlation

Mouse mab 74.5a, by Developmental Studies Hybridoma Bank (2 mentions) Nembutal, by Abbott (2 mentions) Thionin, by Merck Group (2 mentions) Bodioy tmr x muscimol, by Thermo Fisher Scientific (2 mentions) Hydroethidine, by Merck Group (1 mentions) Confocal microscopy, by Zeiss (1 mentions) Dapi solution, by Vector Laboratories (1 mentions) Claudin 5, by Thermo Fisher Scientific (1 mentions) Citrisolv, by Thermo Fisher Scientific (1 mentions) Permount, by Thermo Fisher Scientific (1 mentions) Rabbit pab, by Fujifilm (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Microtome (145 citations) HM 450 Sliding Microtome (13 citations) Microm HM 450 sliding microtome (7 citations) HM430 sliding microtome (5 citations) Freezing sliding microtome (4 citations) Sliding-base freezing microtome (2 citations) MicRoM HM 325 sliding microtome (2 citations) HM450 freezing-sliding microtome (2 citations)

9 protocols using sliding microtome

In Situ Visualization of Oxidants

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hydroethidine histochemistry was performed for in situ visualization of the O₂- and O₂-derived oxidants. Three days after pKr-2 injection, hydroethidine (1 mg/kg in PBS containing 1% dimethylsulfoxide; Sigma, St. Louis, MO, USA) was intravenously administered through tail vein. After 45 min from hydroethidine injection, the brain tissues were prepared, as previously described [40 (link),52 (link)]. The brain tissues were cut into 40 µm using a sliding microtome (Thermo Scientific, Walldorf, Baden-Württemberg, Germany) and the tissues were mounted on gelatin-coated slides. The oxidized hydroethidine product, ethidium, examined by confocal microscopy (Carl Zeiss), and then merged with DAPI solution (Vector Laboratories). Image J quantified the obtained images in each group (National Institutes of Health, USA).

Jeong J.Y., Chung Y.C, & Jin B.K. (2019). Interleukin-4 and Interleukin-13 Exacerbate Neurotoxicity of Prothrombin Kringle-2 in Cortex In Vivo via Oxidative Stress. International Journal of Molecular Sciences, 20(8), 1927.

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Histological Analysis of Brain Tissue

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Animal brains were harvested at either two weeks post-surgery, or following completion of behavioral testing at 2 months, as previously described(16 (link)). The entire brain was sectioned at 50-μm intervals on a sliding microtome (Thermo), and serial sections were mounted, dried, and stained with Cresyl Violet or the following antibodies: BrdU [Rat Mab BU1/75(ICR1), Accurate Chemicals], Olig2 (Mouse Mab from C.D. Stiles, Harvard), GFAP (Rat Mab 2.2B10, Thermo), Iba1 (Rabbit Pab, Wako), CC1 (APC, Mouse Mab OP80, Calbiochem), Nkx2.2 (Mouse Mab 74.5A, Developmental Studies Hybridoma Bank), Collagen 4a (Goat Pab, AB769 Millipore), Claudin 5 (Mouse Mab, 35–2500 Invitrogen).

Nguyen V., Chavali M., Larpthaveesarp A., Kodali S., Gonzalez G., Franklin R.J., Rowitch D.H, & Gonzalez F. (2021). Neuroprotective effects of Sonic hedgehog agonist SAG in a rat model of neonatal stroke. Pediatric Research, 90(6), 1161-1170.

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Perfusion-Fixation and Cresyl Violet Staining

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Immediately following behavioral testing at P38, animals were anesthetized with sodium pentobarbital (100 mg/kg; Nembutal, Abbott Labs, Abbott Park, Ill., USA) and sacrificed. Brains were harvested by transcardiac perfusion with 4% paraformaldehyde (PFA) in 0.1M phosphate-buffered saline (pH 7.4). Brains were carefully removed and postfixed overnight, equilibrated in 30% sucrose in 0.1M PBS and left at 4° C in 0.1M PBS until sectioning and staining. The olfactory bulbs and cerebellum were removed, and the entire brain was sectioned at 50-µm intervals on a sliding microtome (Thermo Scientific, Waltham, MA, USA). The mounted sections were air-dried, stained with cresyl violet, dehydrated in graded ethanol solutions, cleared in Citrisolv (Fisher Scientific, Pittsburgh, PA, USA) and cover slipped in Permount (Fisher Scientific).

Larpthaveesarp A., Georgevits M., Ferriero D.M, & Gonzalez F.F. (2016). Delayed Erythropoietin Therapy Improves Histological and Behavioral Outcomes after Transient Neonatal Stroke. Neurobiology of disease, 93, 57-63.

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Brain Tissue Fixation and Sectioning

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Animals were anesthetized with sodium pentobarbital (100 mg/kg; Nembutal, Abbott Labs) and sacrificed at P63. Brains were harvested by transcardiac perfusion with 4% paraformaldehyde (PFA) in 0.1M PBS (pH 7.4). Brains were removed and post-fixed, equilibrated in 30% sucrose and left at 4° C in 0.1M PBS until sectioning. The entire brain was sectioned at 50-μm intervals on a sliding microtome (Thermo Scientific). Mounted sections were air-dried, stained with cresyl violet, dehydrated, cleared and cover slipped.

Larpthaveesarp A., Pathipati P., Ostrin S., Rajah A., Ferriero D, & Gonzalez F.F. (2020). Enhanced Mesenchymal Stromal Cells or Erythropoietin Provide Long-term Functional Benefit after Neonatal Stroke. Stroke, 52(1), 284-293.

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Brain Tissue Harvesting and Histological Analysis

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Animal brains were harvested at either 2 weeks post-surgery or following completion of behavioral testing at 2 months, as previously described.^{16 (link)} The entire brain was sectioned at 50-µm intervals on a sliding microtome (Thermo), and serial sections were mounted, dried, and stained with Cresyl Violet or the following antibodies: BrdU [Rat Mab BU1/75(ICR1), Accurate Chemicals], Olig2 (Mouse Mab from C.D. Stiles, Harvard), GFAP (Rat Mab 2.2B10, Thermo), Iba1 (Rabbit Pab, Wako), CC1 (APC, Mouse Mab OP80, Calbiochem), Nkx2.2 (Mouse Mab 74.5A, Developmental Studies Hybridoma Bank), Collagen 4A (Col4A; Goat Pab, AB769 Millipore), Claudin 5 (Mouse Mab, 35-2500 Invitrogen).

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Verifying Cannula Placement and Muscimol Diffusion

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Histological verifications of the cannula positions were done after the rats completed all behavioral tests. Rats were anesthetized with sodium pentobarbital and then perfused with ~300 ml PBS and ~300 ml 10% formalin. Brains were removed and stored in 10% formalin and 30% sucrose at 4°C for 3 days before sectioning. Coronal sections (50 μm) of the brain were collected using a sliding microtome (Thermo Fisher Scientific, Waltham, MA). Brain sections were then stained with thionin (Sigma-Aldrich, St. Louis, MO).
To verify the diffusion range of muscimol during the test sessions, the same amount (.25 μl per hemisphere) of fluorescent muscimol (BODIOY TMR-X Muscimol; Molecular Probes, Eugene, OR) was injected before the perfusion. The unstained sections were then examined with a red fluorescent filter. However, it is important to note that the molecular weight of fluorescent muscimol is about six times greater than the muscimol used in this study (Allen et al., 2008 (link)), so the spread of fluorescent muscimol may be underestimated in Figure 2B.

Kim J., Wasserman E.A., Castro L, & Freeman J.H. (2015). Anterior cingulate cortex inactivation impairs rodent visual selective attention and prospective memory. Behavioral neuroscience, 130(1), 75-90.

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Intracellular Labeling and Immunohistochemistry of DRG Neurons

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After electrophysiological characterization and intracellular filling with Neurobiotin, the DRG containing the injected cell was removed and immersion fixed with 3% paraformaldehyde in 0.1 M phosphate buffer (PB) for 30 min at room temperature. Ganglia were then rinsed in 0.1 M PB, embedded in 10% gelatin, postfixed in 3% paraformaldehyde, and cryoprotected in 20% sucrose. 60 μm frozen sections were collected in PB from a sliding microtome (Thermo) and reacted with fluorescently-tagged (FITC) avidin to label Neurobiotin-filled cells (Vector Laboratories). Each section was also processed for ASIC3 (1:2000; Millipore; Cat#: AB5927; RRID: AB_92140) or TRPV1 (1:2000; Alomone; Cat# ACC-030; RRID: AB_2040256) immunoreactivity and isolectin B4 (IB4) binding (AlexaFluor 647; Molecular Probes). After incubation in primary antiserum, tissue was washed and incubated in appropriate fluorescently tagged secondary antibodies (1:200; Jackson Immunoresearch). Distribution of fluorescent staining was determined using Leica confocal microscope. Sequential scanning was performed to prevent bleed-through of the different fluorophores. Images were then captured and compiled using Adobe Photoshop.

Jankowski M.P., Ross J.L., Weber J.D., Lee F.B., Shank A.T, & Hudgins R.C. (2014). Age-dependent sensitization of cutaneous nociceptors during developmental inflammation. Molecular Pain, 10, 34.

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Brain Tissue Harvesting and Processing

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Animal brains were harvested at either two weeks post-surgery, or following completion of behavioral testing at 2 months, as previously described (16) . The entire brain was sectioned at 50-µm intervals on a sliding microtome (Thermo), and serial sections were mounted, dried, and

Nguyen V., Chavali M., Larpthaveesarp A., Kodali S., González G.A., Franklin R.J., Rowitch D.H., & Gonzalez F. (2021). The neuroprotective effects of Sonic hedgehog pathway agonist SAG in a rat model of neonatal stroke.

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Verifying Cannula Placement and Drug Diffusion

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Cannula positions were histologically verified after the rats finished all behavioral tests. The rats were deeply anesthetized with sodium pentobarbital and then perfused with ~300 ml PBS and ~300 ml 10% formalin. Brains were stored in 10% formalin and 30% sucrose at 4˚C for 2 – 3 days before sectioning. Coronal sections (50 μm) of the brain were collected using a sliding microtome (Thermo Fisher Scientific, Waltham, MA) for later thionin staining (Sigma-Aldrich, St. Louis, MO). The diffusion range of the drug was also histologically verified using fluorescent muscimol (BODIOY TMR-X Muscimol; Molecular Probes, Eugene, OR). The same amount (0.3 μl per injection site; 2.0 mM) was infused 30 min before the perfusion. The unstained sections were examined under a red fluorescent filter. It is important to note that the molecular weight of fluorescent muscimol is heavier than the muscimol used for test sessions (Allen et al., 2008 (link)), so the diffusion range may be underestimated in Figure 4.

Kim J., Castro L., Wasserman E.A, & Freeman J.H. (2018). Dorsal hippocampus is necessary for visual categorization in rats. Hippocampus, 28(6), 392-405.

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