Figure 1.
Synthesis of core-shell microspheres.
Reaction conditions: i) PVP 58k, azobisisobutyronitrile (AIBN), hexadecane, 70°C, 16 h ii) AIBN, hexadecane, SDS(aq), RT 70°C, 5 h.
Figure 2.
SEM examination of microspheres.
SEM images of thiouronium-functionalized microspheres 4, core-shell microspheres 6a–6c synthesized with varying ratios of co-monomers to seed particles and core shell microspheres 6e and 6f synthesized with varying ratios of styrene to methacrylic acid.
Table 1.
Composition and sizing data of microspheres 4 and 6a–h.
Table 2.
Zeta potential measurements of microspheres 4 and 6f.
Figure 3.
Fluorescent labeling of core-shell microspheres.
Reaction conditions: i) DMF, MeOH, RT, 16 h; ii) DMF, RT, 2 h; iii) EDAC, MES pH 5.5, DMF, RT, 18 h; iv)EDAC, MES pH 6.5, NaOH, RT, 2.5 h.
Figure 4.
Flow cytometry analysis of microspheres.
Flow cytometry analysis of a) unlabelled microspheres 7a (negative control); b) DY-630 labelled microspheres 8a; c) DY-630 labelled microspheres 8b
Figure 5.
Confocal microscopy of microspheres.
Confocal microscope images of DY-630 labelled microspheres conjugated to GFP 10a: a) excited at 633 nm; b) excited at 488 nm; c) composite image showing co-localization of DY630 and GFP fluorescence. Scale bar = 4 µm.
Figure 6.
Confocal microscopy of a single microsphere.
An overlay confocal microscope image of a DY-630 labelled microsphere conjugated to GFP 10a: excited at 633 nm (core, red) and at 488 nm (shell, green). Scale bar = 2 µm.
Figure 7.
Cellular uptake of core-labeled microspheres, assessed by FACS.
Percentage cellular uptake of DY-630 labelled microspheres 8 by HeLa cells, as measured by flow cytometry: a) 8a (1 µm diameter); b) 8b (500 nm diameter).
Figure 8.
Fluorescent microscopy of beadfected HeLa cells.
Fluorescence microscope images obtained from a Z stack set of images (approximately 0.5 µm z-steps, taken from the top to the bottom focal plane of the cells, 10–12 slices in total) of HeLa cells beadfected with a sample of DY-630-labelled microspheres conjugated via their shells to GFP 10a: a) under irradiation at 633 nm to show DY-630 fluorescence; b) under irradiation at 433 nm to show GFP fluorescence. In each case the cells have been fixed and their nuclei stained with 4',6-diamidino-2-phenylindole (DAPI). Scale bar = 10 µm.
Figure 9.
Ceullular uptake of dual core/shell-labeled microspheres, assessed by FACS.
Cellular uptake of shell-conjugated, DY-630 core-labelled microspheres by HeLa cells, as measured by flow cytometry: a) shell conjugated to fluoresceinamine 9a; b) shell conjugated to GFP 10a.