Celltracker red cmtpx or green cmfda

Linear and branched TEBVs were perfused for seven days to mature. Then, they were treated with 10 μg/ml eLDL and 50 ng/ml TNF-α for four days. On the final two days, monocytes (U937 or THP-1 for linear or branched TEBVs, respectively) were labeled with CellTracker^TM red-CMTPX or green CMFDA (Invitrogen) and perfused through the TEBVs at 1 × 10⁶ monocytes/ml. After two days, the lumens of the TEBVs were washed gently with PBS to remove any unadhered monocytes. TEBVs were then fixed with 4% paraformaldehyde as described in the 3D immunofluorescence staining section. Samples were cut en face and stained with Hoechst dye (1:500) for 2.5 h. Monocyte accumulation was determined by imaging TEBVs and counting the number of cells per area that colocalized cell tracker red and Hoechst dye.
In branched TEBVs, the resulting monocyte adhesion was significantly different at locations, i.e., inlets, main outlets, and side outlets, within a vessel as a result of focal adhesion phenomena characterized in branched arteries. Therefore, monocyte accumulation in branched TEBVs was depicted to indicate the corresponding locations.

Linear and branched TEBVs were perfused for seven days to mature. Then, they were treated with 10 μg/ml eLDL and 50 ng/ml TNF-α for four days. On the final two days, monocytes (U937 or THP-1 for linear or branched TEBVs, respectively) were labeled with CellTracker^TM red-CMTPX or green CMFDA (Invitrogen) and perfused through the TEBVs at 1 × 10⁶ monocytes/ml. After two days, the lumens of the TEBVs were washed gently with PBS to remove any unadhered monocytes. TEBVs were then fixed with 4% paraformaldehyde as described in the 3D immunofluorescence staining section. Samples were cut en face and stained with Hoechst dye (1:500) for 2.5 h. Monocyte accumulation was determined by imaging TEBVs and counting the number of cells per area that colocalized cell tracker red and Hoechst dye.
In branched TEBVs, the resulting monocyte adhesion was significantly different at locations, i.e., inlets, main outlets, and side outlets, within a vessel as a result of focal adhesion phenomena characterized in branched arteries. Therefore, monocyte accumulation in branched TEBVs was depicted to indicate the corresponding locations.