The cerebella of mice older than P21 were considered mature. 1 mm-thick midsagittal mouse brain sections were obtained using a mouse brain matrix (ASI-Instruments, RBM-2000C) and used to make paraffin blocks. Paraffin-embedded cerebellar sections (7 μm-thick) were de-paraffinized via standard procedures and stained with cresyl violet (Sigma-Aldrich, #C5042) or nuclear fast red solution (Sigma-Aldrich, #N3020) for 1–5 min at room temperature. The sections were mounted using Cytoseal™ XYL (Thermo Scientific, #8312–4) and scanned with Aperio Scanscope XT scanner (Aperio Technologies). Cresyl violet-stained images of the whole cerebellar sections and the GCL were obtained, converted to black-and-white, and used to measure the occupancy of the GCL and ML via the ImageJ 1.50 program (NIH) [33 (link)]. The area covered by the ML was measured by subtracting the area covered by the GCL and white matter from the area of the whole cerebellum.
Nuclear fast red solution
Manufactured by Merck Group
Sourced in United States, Germany
Nuclear Fast Red solution is a laboratory reagent used in histology and cytology. It is a staining dye that selectively binds to nucleic acids, enabling the visualization of cell nuclei under a microscope. The solution is typically used in various staining protocols to highlight the presence and distribution of cellular nuclei within tissue samples or cell preparations.
Automatically generated - may contain errors
Lab products found in correlation
Alcian blue solution, by Merck Group (10 mentions)
Bx61 microscope, by Olympus (3 mentions)
Entellan, by Merck Group (3 mentions)
Alcian blue, by Merck Group (3 mentions)
Axio scan z1 slide scanner, by Zeiss (2 mentions)
Iron stain kit, by Merck Group (2 mentions)
Axioskop 40, by Zeiss (2 mentions)
Bx51 microscope, by Olympus (2 mentions)
Depex mounting medium, by Serva Electrophoresis (2 mentions)
Aqueous silver nitrate solution, by Merck Group (2 mentions)
Alizarin red s solution, by Merck Group (2 mentions)
T7 sp6 polymerase synthase kit, by Promega (2 mentions)
Anti digoxigenin ap, by Roche (2 mentions)
Dig rna labelling mix, by Roche (2 mentions)
Nbt bcip, by Roche (2 mentions)
Xylene, by Duksan Pure Chemicals (2 mentions)
Potassium ferrocyanide, by Merck Group (2 mentions)
Alcian blue 8gx solution, by Merck Group (2 mentions)
Sodium thiosulfate, by Merck Group (2 mentions)
Ishyb in situ hybridization kit, by BioChain (2 mentions)
72 protocols using nuclear fast red solution
1
Cerebellar Maturation Analysis in Mice
2
Alcian Blue Staining of Intestinal Goblet Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Alcian Blue staining, the intestinal tissues were fixed in Carnoy’s solution (60% ethanol, 30% chloroform, and 10% glacial acetic acid) for 4 h at room temperature. Subsequently, the tissues were transferred in 100% ethanol overnight and further processed for paraffin sectioning. Following deparaffinization, slides were stained in Alcian blue solution (3%, pH 2.5, Sigma) for 30 min. Slides were washed in tap water for 2 min, counterstained with nuclear fast red solution (Sigma-Aldrich) for 5 min, and followed by dehydration in ethanol gradient solution. Goblet cells were quantified using the Cell counter ImageJ software and normalized to crypt numbers.
3
Quantification of Alcian Blue-Positive Cells in FFPE Lung Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FFPE lung sections were deparaffinized and incubated during 1 h in 1% Alcian Blue 8GX solution in 3% acetic acid (Sigma). A counterstaining was performed using Nuclear Fast Red solution (Sigma) during 15 min. Microscopy was performed using a Nikon DS Fi2 camera and NIS-Elements software (Nikon). Positive cells for Alcian Blue staining (AB+) were manually counted in the main bronchi (Fig. S2 ) and results were expressed as cells/mm epithelium.
4
Histological Analysis of Mouse Organs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All tissue used in these experiments was harvested from mice sacrificed at desired age by intraperitoneal injection of sodium pentobarbital, in accordance with UK the guidance and rules for the use of animals in research and were carried out at the University of Edinburgh. Whole liver and heart were rapidly dissected and fixed for 4 h in 4% paraformaldehyde (PFA). Sequentially, one part of the liver and whole heart were then cryoprotected in 30% sucrose for subsequent OCT embedding, and the other liver part submerged in 70% ethanol to be processed in wax. After sectioning (5 μm), wax embedded liver and OCT embedded heart were stained in Haematoxylin and Eosin (H&E) or for iron (Prussian blue reaction – Mallory’s method) co-labeled with 0.1% Nuclear fast red solution (Sigma-Aldrich N3020) following a standard protocol.
5
Tissue Iron Detection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fixed brain sections were immersed in Perls solution consisting of 2% HCl (v/v) and 2% potassium ferrocyanide (w/v) in a 1:1 ratio for 30 min at room temperature. Sections were washed and counterstained with Nuclear Fast Red solution (Sigma) for 5 min. The sections were dehydrated under alcohol gradient (75, 90 and 100%) followed by two changes of xylene. Finally, the sections were mounted using DPX and viewed with a BX61 microscope (Olympus).
6
Cryosectioning and Histochemical Staining of Utrn Mutant Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues from UtrnR and WT mice were incubated in cold lacZ fixative (2% formaldehyde, 0.2% glutaraldehyde, 0.02% Nonidet P-40, 1 mM MgCl2, 0.1 mg/ml Sodium Deoxycholate in PBS) for 1 h at 4 °C. Samples were incubated in 30 % sucrose for 24 h, embedded in O.C.T compound and stored at −80 °C. Frozen tissue was sectioned to 20 µm thickness using a cryostat and collected onto Superfrost slides. Defrosted slides were stained with lacZ stain (0.5 mg/ml X-Gal, 4 mM Potassium Ferrocyanide, 4 mM Potassium Ferricyanide, 1 mM MgCl2, 0.02% Nonidet P-40 in PBS) at 37 °C for 5 h. After staining, tissue was fixed in 4% PFA for 10 min then washed (1 × 5 min PBS, 1 × 10 min PBS, 2 × 5 min dH2O). Counter staining was performed by 2 min incubation with Nuclear Fast Red solution (Sigma) followed by two 5 min washes in dH2O. Slides were mounted using Prolong Gold Antifade mountant (Thermo Fisher Scientific) and imaged on an Axio Scan.Z1 Slide Scanner (Zeiss).
7
Perls' Stain for Iron Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Renal tissue slides from Sham- and CLP-treated mice were dewaxed in xylene and rehydrated in a series of alcohol solutions using decreasing concentrations. Perls’ stain was performed using the Iron Stain Kit (Sigma Aldrich, Germany) according to the manufacturer’s protocol. Slides were then washed in distilled water, counterstained with nuclear fast red solution (Sigma Aldrich), rapidly dehydrated, and mounted in entellan (Merck). Pictures were acquired using an Axioskop 40 (Zeiss, Wetzlar, Germany).
8
Mineralization Assay: Histochemical Staining
9
Cellular Uptake of Nanoparticles Quantified
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To confirm the cellular uptake of nanoparticles, 1×106 HUVECs were incubated with NGD-MNPs at a concentration of 20 μg/mL for 6 h. The cells were washed with PBS and fixed in 4% paraformaldehyde for 15 min. After removing the fixative, 1 mL of solution containing 5 wt% potassium ferrocyanide and 10 vol% HCl were added to each well and incubated at room temperature for 15 min. After washing with distilled water three times, the cells were counterstained with nuclear fast red solution (Sigma Chemical Co, St. Louis, MO) for 3 min. Images were obtained under Olympus BX51 microscope (Olympus, Tokyo, Japan).
The quantitative analysis of iron content internalized by HUVECs was determined by an atomic absorption spectrophotometer (AAS, Shimadzu Corp, Kyoto, Japan). 1×106 HUVECs were treated with CoCl2 as described above and then incubated with nanoparticles (20 μg/mL) for 6 h. The cells were lysed with 200 μL of concentrated HCl and the lysate was transferred into a micro reaction tube. After an appropriate dilution of lysate with distilled water, intracellular iron content was measured by AAS.
The quantitative analysis of iron content internalized by HUVECs was determined by an atomic absorption spectrophotometer (AAS, Shimadzu Corp, Kyoto, Japan). 1×106 HUVECs were treated with CoCl2 as described above and then incubated with nanoparticles (20 μg/mL) for 6 h. The cells were lysed with 200 μL of concentrated HCl and the lysate was transferred into a micro reaction tube. After an appropriate dilution of lysate with distilled water, intracellular iron content was measured by AAS.
10
Senescence Assay in Growth Plate
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess cellular senescence in the growth plate, freshly dissected growth plates were embedded in OCT and snap-frozen on dry ice and maintained at −20 °C. Frozen sections of growth plate were immediately prepared, fixed for 1 minute in 5% formalin, and incubated overnight at 37 °C in 1 mg/ml of X-gal (5-bromo-4-chloro-3-indolyl β-D-galactopyranoside; Invitrogen), 5 mM K3Fe(CN)6 (Sigma-Aldrich), 5 mM K4Fe(CN)6×3H2O (Sigma), and 40 mM citric acid/sodium phosphate (pH 6.0) as previously described [41 (link)]. The reaction was stopped by washing the slides in PBS (pH 7.4) and fixing in 10% formalin, followed by counterstaining with 0.1% nuclear fast red solution (Sigma), dehydrating in serial ethanol solutions (70%, 85%, and 100%, 5 minutes each), immersing in xylene, and mounting using Permount solution (Fisher Scientific).
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!