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. Author manuscript; available in PMC: 2014 Mar 13.
Published in final edited form as: Int J Cardiol. 2013 Jul 22;168(4):3659–3664. doi: 10.1016/j.ijcard.2013.05.092

Figure 2. Assessment of lipoplex formation and stability by flow cytometry and uptake of Dz13 in DOTAP/DOPE in rabbit veins ex vivo.

Figure 2

Figure 2

Figure 2

(A) To assess the lipoplex stability during material transport and surgical preparation, the Dz13 (500 µg) /DOTAP/DOPE mixture were incubated at 22°C for 10 minutes, followed by incubation on ice (0°C) for 0.5, 1, 2 and 3 hours. The FSC/SSC scatter plot of the lipoplexes at different time points were recorded by flow cytometry. (B) Using flow cytometry, similarly, to assess the lipoplex stability during incubation with the veins, the Dz13 (500 µg) /DOTAP/DOPE mixture were incubated at 22°C for 10 minutes and on ice (0°C) for 1 hour, followed by incubation at 37°C for up to 3 hours. (C) Rabbit jugular veins were incubated in 300µl transfection solution containing 500µg FAM-Dz13 and 37.5µl DOTAP/DOPE at 37°C for 30 min (after incubation of the Dz13/DOTAP/DOPE mixture for 10 min at 22°C, and 1h at 0°C) then rinsed in PBS and snap frozen. Blocks were sectioned, placed onto slides and fixed in 95% ethanol. DAPI nuclear staining (blue) was imaged under fluorescence microscopy. Green fluorescence demonstrates that Dz13 was delivered into rabbit veins.