An immunosorbent assay system was integrated into a glass microchip. Polystyrene beads were introduced into a microchannel, and then human secretory immunoglobulin A (s-IgA) adsorbed on the bead surface was reacted with colloidal gold conjugated anti-s-IgA antibody and detected by a thermal lens microscope. The scale merits of liquid microspace on the molecular behavior remarkably contributed to reduced assay time. The integration cut the time necessary for the antigen-antibody reaction by 1/90, thus shortening the overall analysis time from 24 h to less than 1 h. Moreover, troublesome operations required for conventional immunosorbent assays could be replaced by simple operations.

We have fabricated nanometer-sized channels, demonstrated a technique for the introduction of liquid into the channels, and carried out time-resolved fluorescence measurements of aqueous solutions. In this study, 330-nm- and 850-nm-sized channels were fabricated on fused-silica substrates by fast atom beam etching and hydrofluoric acid bonding methods. A liquid introduction method utilizing capillary action was demonstrated. The liquid introduction was observed under an optical microscope, and the liquid velocity during the introduction was analyzed by surface energy and macroscale hydrodynamics. The liquid velocity due to capillary action in the nanometer-sized channel seemed more than four times slower than the estimation. Then, aqueous solutions of rhodamine 6G (R6G), sulforhodamine 101 (SR101), and rhodamine B (RB) in the channels were measured by time-resolved fluorescence spectroscopy; spectra of the same solution in a 250-microm-sized channel were also measured as a reference for the macrospace. Although the fluorescence spectra in the 330-nm-, 850-nm- and 250-microm-sized channels agreed with one another, the fluorescent decays in the nanometer-sized channels were faster for R6G and SR101 and slower for RB than the respective decays in the 250-microm-sized channels. The results suggested the solutions had lower dielectric constants and higher viscosities in the nanometer-sized channels.

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