Application of total internal reflection fluorescence microscopy to study cell adhesion to biomaterials

Department

Bioengineering

Document Type

Article

Publication Title

Biomaterials

ISSN

0142-9612

Volume

19

Issue

4--5

DOI

10.1016/S0142-9612(97)00109-9

First Page

307

Last Page

325

Publication Date

3-1-1998

Abstract

Cell adhesion and function depend upon the formation of adhesive contacts between the cell and substrate. Determination of the cell substrate contact area is necessary in order to understand how biomaterial properties influence cell adhesion. In this review we describe the development and application of total internal reflection fluorescence microscopy (TIRFM) to quantify the separation distance of cells from a biomaterial surface. An approximate theory is presented for the straightforward calculation of separation distances when a fluor is placed in the cell membrane. The validity of this approach is discussed. TIRFM is compared to interference reflection microscopy and related techniques that measure cell/substrate separation distances. This approach is then applied to a number of important problems in cell substrate interactions, including changes in contact area and adhesion strength on biomaterial surfaces, analysis of bond strength, and real-time measurement of cell/substrate separation distances following exposure to flow.

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