All surgical procedures were performed under aseptic conditions in a designated animal surgery area. Mice were anesthetized by inhaled isoflurane (4% induction, 2.5 % maintenance) and using a Hamilton microsyringe (33 gauge needle), multiple 1.5-2µl injections of Alexa Fluor 488-conjugated cholera toxin-β (CTB, 2mg/ml in saline, 8µl total; Invitrogen) were made circumferentially within the distal region of the vagina, approximately 3–5mm from the external opening. Using a separate Hamilton microsyringe (33 gauge needle), 2–3µl injections of 1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI; 2% in DMSO, 5µl total; Sigma, St. Louis, MO) were made on the right and left side, subcutaneously, at the junction between the hairy skin and the opening of the vagina. Mice were returned to their home cages and allowed to recover from anesthesia under observation.
Tetramethylindocarbocyanine perchlorate dii
Manufactured by Merck Group
1,1'-Dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) is a fluorescent dye commonly used in biological research. It is a lipophilic carbocyanine dye that incorporates into cell membranes and is useful for cell labeling and tracing applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using tetramethylindocarbocyanine perchlorate dii
1
Vaginal Tract Labeling in Mice
2
Lipid-associated APOA1 Vesicle Formation
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Lipid-associated APOA1 were generated following previous protocols with slight modifications (54 (link), 55 (link)). Briefly, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) (Avanti Polar Lipids), with or without 1,1’ -Dioctadecyl-3,3,3′,3′-Tetramethylindocarbocyanine Perchlorate (DiI) (Sigma), were transferred to a 5 ml or 10 ml round bottom flasks manufactured from borosilicate 3.3 glass with a 14/20 Joint size (VWR) and dried to a film by streaming N2. This film was resuspended in methanol and dried to film by N2 streaming. The film was resuspended in 25 mM Hepes in PBS and sonicated at 24°C for 20 min or until the lipid film was fully resuspended. Recombinant APOA1 equilibrated to 24°C was added to the resuspended lipids and further sonicated for 30 min at 24°C or until turbidity disappeared. This solution was incubated at 24°C overnight to allow APOA1 incorporation into vesicles.
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