After fixation with PFA, spleens were frozen-embedded in optimal cutting temperature (O.C.T.) compound (Tissue-Tek, VWR). 10 μm thick sections were obtained with a cryostat microtome (Microm HM 505e) and transferred to charged microscope slides (VWR). Slides were stored at –80° C, then thawed in PBS at room temperature, washed 3x in PBS, and stained with Hoechst (1:10,000 dilution in PBS) for 5 minutes. Slides were then washed 3x with PBS and coverslips were mounted with ProLong Gold (Life Technologies). One section was imaged for each tissue. Tissues were imaged with a Zeiss Axio Observer 7 (Zeiss) inverted fluorescent microscope with an Apotome.2 (Zeiss) with the 63x oil immersion objective. Images of the sections were captured with an Axiocam 702 mono camera (Zeiss).
Charged microscope slides
Manufactured by Avantor
Charged microscope slides are laboratory equipment used to mount and secure samples for microscopic examination. These slides have a surface charge that helps adhere and hold samples in place during the observation process. The charged surface allows for secure positioning of the sample without the need for additional adhesives or coatings.
Automatically generated - may contain errors
2 protocols using charged microscope slides
1
Cryosectioning and Immunofluorescence of Spleen
2
Cryopreservation of Organoid Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Organoids were fixed in 4% PFA overnight at 4°C. Organoids were then transferred to 15% sucrose and 0.05% sodium azide in PBS for 24 h, followed by another 24 h in 30% sucrose and 0.05% sodium azide in PBS. Organoids were then embedded in OCT freezing medium and snap frozen on an ethanol dry ice slurry. Frozen embedded organoids were given at least 3 h to warm to −20°C before cryosectioning on a Leica cryostat at thicknesses between 10 and 30 μM. Sections were mounted on charged microscope slides (VWR) and placed on a slide warmer at 37°C for 30 min. Slides were then stored in a slide box at −20°C until used for staining.
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!