Evanescent Detection of Adsorbed Protein Concentration-Distance Profiles: Fit of Simple Models to Variable-Angle Total Internal Reflection Fluorescence Data

Authors: Reichert, W. M.; Suci, P. A.; Ives, J. T.; Andrade, J. D.

Source: Applied Spectroscopy, Volume 41, Issue 3, Pages 349-531 (March/April 1987) , pp. 503-508(6)

Publisher: Society for Applied Spectroscopy

Buy & download fulltext article:

OR

Price: $29.00 plus tax (Refund Policy)

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.

Keywords: ATR spectroscopy; Analysis for protein adsorption; Fluorescence; Surface analysis

Document Type: Research Article

DOI: http://dx.doi.org/10.1366/0003702874448913

Affiliations: Department of Bioengineering, and Center for Biopolymers at Interfaces, University of Utah, Salt Lake City, Utah 84112

Publication date: March 1, 1987

More about this publication?
Related content

Tools

Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content

Text size:

A | A | A | A
Share this item with others: These icons link to social bookmarking sites where readers can share and discover new web pages. print icon Print this page