Lipid Labeling for Cellular Imaging

Fatty acids were from TCI America (Portland, OR, USA), PEG was from Laysan Bio (Arab, AL, USA), and MTz-NHS was from Click Chemistry Tools. DiI-C18:2, and DiI-C16, DiI-C12 were from ThermoFisher (Waltham, MA, USA). DiI-C18 (1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindotricarbocyanine Perchlorate) and DiR (1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindotricarbocyanine Iodide) were from Biotium (Hayward, CA, USA). DiI-NH₂ was synthesized by a previously reported method.^{[10 (link)]} All the lipids were stored as 1 mM stocks in ethanol. Anticoagulant Citrate Dextrose (ACD) buffer was obtained from the Children’s Hospital Colorado Blood Donation Center and kept sterile before use. Goat IgG fraction to mouse C3 and human C3 (Catalog No. 55 463 and 55 033, respectively) was from MP Biomedicals (Solon, OH, USA). ChromPure human and mouse IgG was from Jackson Immuno Research (West Grove, PA, USA). Secondary antibodies IRDye 80°CW labeled donkey antigoat, goat antihuman, and goat antimouse IgG were from LI-COR Biosciences (Lincoln, NE, USA).