Retroobital injection of BV13-AF647.

Video legend
After putting the mouse in the isoflurane chamber for 3 min, the mice become anesthetized (not shown in video). After confirming the state of anesthesia by pinching the in-between-toe area and lack of movement of mouse, the mouse was taken out from the isoflurane chamber and subjected to retro-orbital injection. Position the head of mouse closer to the needle. Firmly hold the eye area by placing the thumb and index fingers at each side of the eye ball. Using a downward motion, expose the eye ball as much as possible and meanwhile maintaining the eyeball in still position (This required placement of thumb and index fingers immediately adjacent of eye ball, but not further down below). Insert the needle from the anterior end of the eye to the area subneath the eyeball, (optional: one can inject the needle fully until the needle feels the bones behind the retroorbital plexus and retrieve the needle a little bit). Inject the 25 μg of BV13-AF647 diluted in 75 μl PBS (in total 100 μl volume) into the mouse. Quickly detach the needle and gently cover the eyes with eye lids to prevent bleeding. Mice were monitored for any signs of bleeding or other discomfort, which usually do not take place. (Study approval. All animal experiments were performed under the approval of Weill Cornell Medicine Institutional Animal Care and Use Committee, New York, NY. All experimental procedures followed the IACUC guidelines. This video was made at Weill Cornell Medical College according to the guidelines of the IACUC of Weill Cornell Medical College, New York, New York, USA under protocol # 2009-0061.)

Extracted from protocol
Generation of BMEC Lines and in vitro BMEC-HSPC Co-culture Assays
Guo, P. and Rafii, S. (2018). Bio-protocol 8(21): e3079.
DOI: 10.21769/BioProtoc.3079.