All procedures involving animals were approved by and conformed to the guidelines of the West China Hospital Animal Care Committee. We have made great efforts to reduce the number of animals used in these studies and also made efforts to reduce animals suffering from pain and discomfort. The brain tissue was isolated from P1, P7, P15, P20, P90 Specific pathogen Free (SPF) Kumming (KM) mice after euthanasia. Tissues were put into 4% paraformaldehyde for 3–5 days and then dehydrated in 30% sucrose solution. The fixed mouse brain slices (20 μm) were obtained by Leica CM3050S freezing microtome. The slices were stained with complex MyL-1 (20 μM) for 30 min at room temperature. The imaging was performed after the slices were washed by PBS 3 times. To avoid misclassification due to individual differences in animals, at least three independent replicates were set up for each animal experiment.
Cm3050s freezing microtome
Manufactured by Leica
The CM3050S is a freezing microtome from Leica, designed for sectioning frozen tissue samples. It features a cryogenic chamber that maintains a low temperature to preserve the specimen during the cutting process. The microtome allows for precise and consistent sectioning of samples, enabling accurate analysis and examination.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using cm3050s freezing microtome
1
MyL-1 Staining of Mouse Brain Slices
2
TUNEL Assay on Mouse Eye Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wild type (WT) C57/bl6 mice were sacrificed at 5 weeks. Eyes were mounted in Tissue Tek O.C.T. and frozen with liquid nitrogen immediately after enucleation. Then, 10 μm frozen sections were cut on a Leica CM3050S freezing microtome. Sections were incubated in 4% formaldehyde in PBS for 15 min and in 0.3% Triton X-100/PBS for 15 min. Enzymatic digestion with either DNase I or DNase II were then performed. For DNase I digestion, sections were incubated 30 min at 37°C with 2 units of DNase I in associated buffer (#EN0521, Thermo Scientific). For DNase II digestion, sections were incubated 30 min at 37°C with 10 units of L-DNase II (Worthington) in 120 mM Tris-HCl, 120 mM EDTA buffer (pH 5,5). Some sections were then dephosphorylated with 10 units of calf intestinal alkaline phosphatase (CIAP) in the associated buffer (Invitrogen, 8009-019) for 30 min. Finally, the TUNEL assay was performed on all sections following the manufacturer's instructions (Roche Diagnostics, Mannheim, Germany).
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!