The absolute brightness of fluorescent particles, such as dye-containing nano- and microspheres or quantum dots, is a critical design parameter for many applications relying on fluorescence detection. The absolute brightness, defined as the ratio of radiant intensity of emission to illumination intensity of excitation, of nile-red fluorescent microspheres with a 1 micrometre diameter is measured to be 4.2 +/- 1 x 10(-16) m(2)/sr, and the implications for the design of kinesin motor protein-powered "smart dust" devices and the remote detection of fluorescence are discussed.