Whole brains were dissected between E13.5 and E18.5 and fixed for at least 48 h with 4 % PFA in PBS at 4 °C. After fixation brains were washed with PBS. For thalamic injection the brains were cut in half at the midline in the sagittal plane and a small hole was made in the medial aspect of the thalamus using a fine probe. Crystals of DiI (Invitrogen) were inserted into the prepared hole in the thalamus. For cortical or ventral telencephalic injection, holes were made in the desired region of the telencephalon without any further dissection of the brain and crystals of either DiI or DiA were inserted. Brains were incubated in PBS at 37 °C. E14.5 brains were incubated for 1 week while E16.5 and E18.5 brains were incubated for 3–4 weeks. After diffusion the brains were embedded in agarose and sectioned either coronally (telencephalic injections) or at a 45° angle (thalamic injections) at 100 μm. Sections were counterstained with TOPRO-3 diluted 1/1000 in PBS.
Crystals of dii
Manufactured by Thermo Fisher Scientific
Crystals of DiI are a type of lipophilic dye used in various biological applications. The dye can be incorporated into cell membranes, allowing for the visualization and tracking of cells in research and experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using crystals of dii
1
Dye Tracing of Developing Mouse Brain
2
Retrograde Labeling of Cerebellar Granule Cell Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After perfusion with 4% PFA, the cerebellum was dissected and postfixed by immersion in 4% PFA at least overnight. Crystals of DiI (D3911, Invitrogen) were inserted on the surface of the cerebellum in the ML to target the axons of the GCNs so their cell bodies could be retrogradely labeled. Crystals were kept in place with 6% agarose. The cerebellums were then placed in 4% PFA at 37°C for 3 to 7 days. Cerebellums were sectioned (30 μm) using a vibratome (Leica VT1200S) and placed onto slides to adhere before mounting with Mowiol. Images were obtained on a Zeiss Axioskop microscope (451485 El-einsatz) using a SpotFlex camera driven by Spot software (Spot Imaging Solutions version 4.6.1.41).
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!