Animals were sacrificed using CO2. Mice were transcardially perfused with 100ml phosphatebuffered saline (PBS, pH 7.4) followed by 100ml 4% paraformaldehyde (PFA, pH 7.4, Roth, Cat# 0335) at a rate of 20 ml/min. Brains were post fixed in 4% PFA for 3h at room temperature and were subsequently transferred to a 30% sucrose solution. Coronal brain sections were produced using a sliding microtome (Leica Microsystems, Wetzlar, Germany).
Sliding microtome
Manufactured by Leica Microsystems
Sourced in Germany
The sliding microtome is a precision instrument used to obtain thin, uniform sections of biological samples for microscopic examination. It features a sliding knife that moves horizontally across the sample, allowing for the precise cutting of sections with thicknesses ranging from a few micrometers to several tens of micrometers.
Automatically generated - may contain errors
Lab products found in correlation
Colour view iiiu digital dp27 camera, by Olympus (2 mentions)
Bx50 microscope, by Olympus (2 mentions)
Mi crofrost plus slides, by Thermo Fisher Scientific (2 mentions)
Dpx mounting medium, by Electron Microscopy Sciences (2 mentions)
Lsm 510, by Zeiss (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Sucrose, by Thermo Fisher Scientific (1 mentions)
Sandimmune, by Novartis (1 mentions)
Goat anti il 6, by R&D Systems (1 mentions)
Leica tcs confocal microscope, by Leica Microsystems (1 mentions)
Rabbit anti gfap, by Agilent Technologies (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Axiophot fluorescence microscope, by Zeiss (1 mentions)
Alexa fluor 594 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Lsm 510 confocal microscope, by Zeiss (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Alexa fluor 488 donkey anti rabbit, by Thermo Fisher Scientific (1 mentions)
Rabbit anti gfap, by Merck Group (1 mentions)
Donkey anti chicken alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Mouse anti parvalbumin, by Swant (1 mentions)
26 protocols using sliding microtome
1
Tissue Fixation and Sectioning
2
Xenograft Transplantation in Rats with Immunosuppression
3
Immunohistochemical Analysis of Spinal Cord
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anesthetized and perfused with 4% paraformaldehyde (PFA) in PBS. Spinal cords were removed, separated into portions as described above, and post-fixed in 4% PFA for additional 24 h. Tissues were cryo-protected in 20% glycerol for 24 h before 30-μm free-floating sections were prepared with a sliding microtome (Leica Microsystems, Wetzlar, Germany). Double immunolabeling was performed as previously described [35 (link)]. Briefly, sections were microwaved in 0.01 M citrate buffer (pH 6.0) followed by pretreatment with 1% Triton X in PBS. Sections were blocked with PBS containing 3% normal goat serum and 0.01% Triton X for 30 min and incubated with primary Ab diluted in blocking reagent overnight at 4°C (rabbit anti-GFAP, Dako, Carpenteria, CA, USA, 1:1,000; mouse anti-Iba-1, CCF Hybridoma Core, Cleveland, OH, USA, 1:250; goat anti-IL-6, R&D Systems, Minneapolis, MN, USA, 1:20). To confirm specificity, primary Abs were omitted on adjacent sections. The sections were washed and incubated with species-specific secondary Abs conjugated to FITC or Cy5 (Jackson ImmunoResearch, West Grove, PA, USA) for 2 h at RT. Sections were rinsed, mounted with VECTASHIELD (Vector Labs, Burlingame, CA, USA) and examined on a Leica TCS confocal microscope (Leica Microsystems). Images were analyzed offline with Volocity software version 6.1.2 (PerkinElmer, Waltham, MA, USA).
4
Rat Brain Tissue Sampling for Molecular and Histochemical Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Western blotting, ELISA and qRT-PCR (n = 6 per group for each experiment), rats were anesthetized by i.p. injection of 10% chloral hydrate (400 mg/kg) and sodium pentobarbital (100 mg/kg), brains were rapidly removed from the skull. The hippocampus was quickly dissected on ice and stored at − 80 °C for later analysis. For histochemical experiments (n = 6 per group), rats were perfused transcardially with 4% paraformaldehyde and brain tissues were removed and post-fixed overnight at 4 °C. Tissue samples were embedded with optimal cutting temperature compound. Coronal sections were obtained at the bregma level from − 2.5 to − 3.8 mm, cut at a thickness of 20 μm (1‐in‐6 series, 120 μm apart from each other) with a sliding microtome (Leica Instruments, Germany), and stored at − 20 °C.
5
Transcardial Perfusion and Brain Sectioning
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Animals were sacrificed using CO2. Mice were transcardially perfused with 50 ml phosphate-buffered saline (PBS, pH 7.4) followed by 100 ml 4% paraformaldehyde (PFA) at a rate of 10 ml/min. Brains were post-fixed in 4% PFA for 12 h at 4°C and were subsequently transferred to a 30% sucrose solution. Coronal brain sections were produced using a sliding microtome (Leica Microsystems, Wetzlar, Germany) for phenotyping and morphological analysis.
6
Histological Analysis of Testicular Parasites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Routine paraffin wax embedding procedures were used. Tissue samples from testicular parenchyma, pampiniform plexus and scrotal skin were fixed in 10% buffered formaldehyde, dehydrated in a graded ethanol series, and cleared in xylene. The samples were embedded in paraffin wax, and sections with a thickness of 5 μm were cut using a sliding microtome (Leica Microsystems, Germany). Sections were stained with haematoxylin–eosin (H&E), periodic acid-Schiff (PAS) and Masson trichrome (for better visualization of connective tissue) and underwent a light microscopy evaluation. Photomicrographs of each studied specimen were subjected to computer-assisted image analysis using a computer coupled to an optical Olympus BX50 microscope equipped with a Colour View IIIu digital Olympus DP27 camera (Olympus, Japan). Parasite cysts found in the histological sections were counted in 10 randomized fields with a 10 × magnification objective to obtain an average number of parasite cysts and to measure the diameter of the cysts.
7
Decalcification and Histopathological Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After fixation, the specimens were decalcified with a 10% EDTA (pH 7.4, Ling Feng Chemical Reagent Co., LTD, Shanghai, China) decalcifying solution at room temperature for 4 weeks. After decalcification, the glass ionomer cement and resin composite was carefully removed from the cavity and rinsed with phosphate-buffered saline (PBS, pH 7.4) three times. The specimens were then dehydrated in ascending grades of ethanol, dealcoholized by xylene, and embedded in paraffin. Serial sections of 5 μm thickness were cut using a sliding microtome (Leica Microsystems, Vertrieb, Wetzlar, Germany) for histopathological and immunohistochemical examination [21 (link)].
8
Tissue Preparation for Microscopy
9
Histological Analysis of Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the above procedure, two animals from each group were anaesthetized and submitted to treatment with 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4) by transcardial perfusion. The lung was removed and immersed in the same fixative solution. The lung specimens were sliced into 5 mm pieces, dehydrated through an ascending ethanol series (70, 95, and 100% ethanol for 30 min each) and then embedded in paraffin using standard procedures. Serial 5 µm-thick sections were prepared using a sliding microtome (Leica Microsystems, Wetzlar, Germany). Hematoxylin and eosin (HE) staining wereperformed to quantify possible foci of inflammatory infiltrates and fibrosis in experimental and control animals. The stained sections were observed, and digital images were taken with a Zeiss AxioPhot fluorescence microscope (Carl-Zeiss, Oberkochen, Germany). Specifically, two slices of each sample (six fields of each slice corresponding to approximately 90% of the slice) were used for quantitative analysis to obtain the mean value.
10
Brain Tissue Fixation and Sectioning
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For brain tissue collection, mice were anesthetized using CO2 and transcardially perfused with phosphate‐buffered saline (PBS, pH 7.4) for 5 min at a rate of flow of 20 ml/min followed by fixation with 4% paraformaldehyde (PFA) in 0.1 mM phosphate buffer (pH 7.4, Roth, Cat# 0335) for 5 min at a rate of 20 ml/min. Brains were post‐fixed in 4% PFA at 4°C overnight and subsequently dehydrated in 30% sucrose solution. Frozen brains were either coronally or sagittally cut using a sliding microtome (Leica Microsystems, Wetzlar, Germany). Sections were stored at −20°C in 96‐well plates, filled with 200 μl cryoprotection solution per well.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!