Confocal microscopy of microspheres.

<p>Confocal microscope images of DY-630 labelled microspheres conjugated to GFP <b>10a</b>: 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.</p

Flow cytometry analysis of microspheres.

<p>Flow cytometry analysis of a) unlabelled microspheres <b>7a</b> (negative control); b) DY-630 labelled microspheres <b>8a</b>; c) DY-630 labelled microspheres <b>8b</b></p

Ceullular uptake of dual core/shell-labeled microspheres, assessed by FACS.

<p>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.</p

Cellular uptake of core-labeled microspheres, assessed by FACS.

<p>Percentage cellular uptake of DY-630 labelled microspheres <b>8</b> by HeLa cells, as measured by flow cytometry: a) <b>8a</b> (1 µm diameter); b) <b>8b</b> (500 nm diameter).</p

Fluorescent microscopy of beadfected HeLa cells.

<p>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.</p

Composition and sizing data of microspheres <b>4</b> and <b>6a</b>–<b>h.</b>

a<p>ratio determined on the basis of mass</p><p>Mean diameter and standard deviation of thiouronium-functionalized microspheres <b>4</b> and <b>4a</b> and core-shell microspheres <b>6a</b>–<b>h</b>, as measured dispersed in water using a laser diffractometer.</p

Confocal microscopy of a single microsphere.

<p>An overlay confocal microscope image of a DY-630 labelled microsphere conjugated to GFP <b>10a</b>: excited at 633 nm (core, red) and at 488 nm (shell, green). Scale bar = 2 µm.</p

Fluorescent labeling of core-shell microspheres.

<p>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.</p

Synthesis of core-shell microspheres.

<p>Reaction conditions: i) PVP 58k, azobis<i>iso</i>butyronitrile (AIBN), hexadecane, 70°C, 16 h ii) AIBN, hexadecane, SDS(aq), RT 70°C, 5 h.</p