Abstract

Total internal reflection fluorescence spectroscopy (TIRF) is an established technique for following the course of interfacial reactions. Theoretically, by gathering TIRF data as a function of observation angle, one can obtain the density of fluorophores with respect to distance away from a solid/liquid interface. In order that the practical application of the theory might be explored, variable observation angle data from solutions of fluorescein and from adsorbed layers of fluorescein isothiocyanate labeled immunoglobulin have been analyzed in terms of simple concentration-distance profiles. In all cases the general shape of the data curves was found to conform to the theoretical expectation. Layer thickness determinations varied over a range of 20 to 100 nm, with concentrations in the layer ranging from 12 to 61 mg/mL. The theoretical background, sources of error, and system improvements are also discussed.

PDF Article

Cited By

You do not have subscription access to this journal. Cited by links are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access OSA Member Subscription