This paper focuses on encoding polystyrene microbeads, 10-100 microm in diameter, with a luminescent spectral bar code composed of mixtures of quantum dots (QDs) emitting at different wavelengths (colors). The QDs are encapsulated in the bead interior during the bead synthesis using a suspension polymerization, and the bar code is constructed by varying both the number of colors included in the bead and, for each color, the number of QDs of that color. Confocal laser scanning microscopy images of the beads demonstrate that the multicolored QDs are pushed together into inclusions within the bead interior. The encoded bead emission spectrum indicates that the peak position of the included colors does not shift relative to the corresponding peaks of the spectra recorded for the nonaggregated QDs at identical loading concentrations. Due to the spatial proximity of the QDs in the inclusions, electronic energy transfer from the lower wavelength emitting QDs to the higher emitting QDs changes the relative intensities of the colors compared to the values in the nonaggregated spectra. We show that this energy transfer does not obscure the spectral uniqueness of the different codes. Ratiometric encoding, in which the bar code is read as relative color intensity, is shown to remove the dependence of the code on the bead size.

Here, we describe a protocol to bind individual, intact phospholipid bilayer liposomes, which are on the order of 1 microm in diameter, in microwells etched in a regular array on a silicon oxide substrate. The diameter of the wells is on the order of the liposome diameter, so only one liposome is located in each well. The background of the silicon oxide surface is functionalized with a PEG oligomer using the contact printing of a PEG silane to present a surface that resists the adsorption of proteins, lipid material, and liposomes. The interiors of the wells are functionalized with an aminosilane to facilitate the conjugation of biotin, which is then bound to Neutravidin. The avidin-coated well interiors bind the liposomes whose surfaces contain biotinylated lipids. The specific binding of the liposomes to the surface using the biotin-avidin linkage, together with the resistant nature of the background and the physical confinement of the wells, allows the liposomes to remain intact and to not unravel, rupture, and fuse onto the surface. We demonstrate this intact arraying using confocal laser scanning microscopy of fluorophores specifically tagging the microwells, the lipid bilayer, and the aqueous interior of the liposome.