The whole right hind-limb was resected during [18F]-FDG biodistribution. Tissue was incubated at 4 °C for 1 h in optimal cutting temperature media (Fisher Healthcare, Houston, TX, USA). The limb was then transferred to a fresh optimal cutting temperature-filled mold and flash frozen on dry ice. The limb was rapidly sectioned using a custom modified CM1800 cryostat (Leica, Wetzler, Germany) at 10 μm. Standard sensitivity phosphor-screens (PerkinElmer) were exposed to the section in order to visualize [18F]-FDG uptake as needed for digital autoradiography and read using a Cyclone Phosphorimager (PerkinElmer).
Optimal cutting temperature media
Manufactured by Thermo Fisher Scientific
Sourced in United States
Optimal cutting temperature (OCT) media is a specialized compound used in the preparation of tissue samples for cryosectioning. It provides a stable and supportive matrix to hold the tissue in place during the freezing and sectioning process, allowing for the creation of thin, uniform tissue slices for microscopic analysis.
Automatically generated - may contain errors
4 protocols using optimal cutting temperature media
1
Biodistribution of [18F]-FDG in Tissue
2
Retinal Flat Mount and Sectioning
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rats were euthanized by isoflurane inhalation followed by cervical dislocation which is approved by the Panel on Euthanasia of the American Veterinary Medical Association. Eyes were removed and fixed in 4% paraformaldehyde. Eyes used for flat mounts had the anterior segment and vitreous removed and then retina and the remaining eyecup were isolated and flat mounted separately. Only the RPE is visualized on the surface of the flat mounted eyecup, so this will be referred to as RPE flat mount. Eyes for ocular sections were frozen in optimal cutting temperature media (Fisher Scientific, Walkersville, MD) and 10 μm frozen sections were placed on slides. Sections were stained with Hoechst (Vector Laboratories, Burlingame, CA) and anti-GFP antibody (A21312, life Technologies Corporation, Eugene, OR). Both flat mount and ocular sections were examined by fluorescence microscopy. Two investigators participated in all experiments. One investigator coded slides and the other examined them and obtained images. After the images were obtained the investigators broke the code together to complete the analysis.
3
Retinal Flat Mount and Sectioning
4
Histological Analysis of Animal Eyes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At the end of final IPTG administration, animals were imaged and sacrificed. Eyes were collected and fixed in 4% paraformaldehyde. Tissues were then cryoprotected and embedded in optimal cutting temperature media (Fisher Scientific Co., Pittsburgh, PA, USA) and frozen. Cryosections were made using a Leica CM1850 cryostat (Leica Microsystems, Wetzlar, Germany). Sections were then stained with hematoxylin and eosin (H&E).
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!