Extracellular vesicles labeling and tracking in vitro and in vivo

CY Chaoqun Yu
PC Peng Chen
JX Jing Xu
YL Yaning Liu
HL Hui Li
LW Linna Wang
GD Guohu Di
request Request a Protocol
ask Ask a question
Favorite

To detect the direct transfer of extracellular vesicles into HCECs, hADSC-Evs were labeled using PKH67 Fluorescent Cell Linker Kit (PKH67) (Cat#MINI67-1KT, Sigma, Saint Louis, USA), and then incubated with HCECs for 4 h, followed by fixation and (imaging. To evaluate the uptake of extracellular vesicles in vivo, normal female mice were topically applied with PKH67-labled hADSC-Evs (1 μg/μL) with 4 times a day (8:00 am; 11:00 am; 14:00 pm; 17:00 pm; n = 4; 5 μL/eye/time). 2 h after the last treatment, eyeballs were fully rinsed by PBS and snap frozen in Tissue-Tekoptimum cutting temperature compound. Immunofluorescent staining of frozen corneal section (7 μm thick) was performed and observed under microscope (Nikon).

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A