DETECTING BIOMOLECULES IN PICOLITER VIALS USING AEQUORIN BIOLUMINESCENCE

The quantitative determination of proteins in picoliter-volume vials i s described. The assay is based on the bioluminescence of the photopro tein aequorin along with photon-counting detection. Using this approac h, avidin can be detected at femtomole levels by taking advantage of i ts inhibitory effect on the bioluminescence signal generated by biotin ylated recombinant aequorin. The picoliter vials were fabricated on gl ass substrates using a laser ablation technique. Parameters that affec t the reproducibility of the assay such as the fabrication and calibra tion of the pipets, the fabrication of the vials, and the composition of the assay solutions were studied.