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. 2023 Apr 19;66(9):6288–6296. doi: 10.1021/acs.jmedchem.3c00138

Figure 4.

Figure 4

Alkyne-tagging photolinker and permeation assay in microfluidic droplets. (A) Reductive amination of aldehyde photocleavable (PC) linker with PPA yielded the alkyne-tagging photolinker (PC–PPA). Subsequent acylation with various acids yielded photocleavable alkyne-tagged permeation control beads (PC–PPA–V/PC–PPA–PEG17). Alkynes were released from the bead for permeation assay upon UV irradiation. Control beads (B) PC–PPA–V and (C) PC–PPA–PEG17 were evaluated in droplet-scale liposomal permeation assays. The photocleaved product, PPA–V, is a permeable positive control that increases fluorescence of the liposomal probe; PPA–PEG17 an impermeable negative control that does not increase fluorescence of the liposomal probe. Transient heat maps display the ouput from microfluidic droplet-scale permeation assays of PC–PPA–V and PC–PEG17 control beads. A calculated threshold 5 standard deviations (5σ) above the mean negative signal (green line) was used to calculate the false discovery rate. The droplet generation rate (gray), bead occupancy rate (red), and hit rate (black) were binned for the PC–PPA–V and PC–PEG17 microfluidic analyses in 30 s windows.