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Proceedings Paper

Measuring the surface topography at focal adhesions using laser feedback interferometry
Author(s): Michael Bambino; Ben Ovryn
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Paper Abstract

Reflection interference contrast microscopy of living cells has not yet fully matured. One goal would be to temporally resolve the distance between the cell and the substratum at each point over the cell surface. We have combined phase shifting laser feedback interferometry with a high numerical aperture inverted microscope in order to determine the topography of the ventral surface of a cell. We have obtained a map of both the topography of a cell as well as its reflectivity. Our data demonstrate that interference microscopy can be adapted to yield a measure of the distance between the cell and the substratum. We have quantified the ventral surface topology at focal adhesions and we have shown that these changes are correlated with markers for a focal adhesion adaptor protein. The laser feedback interferometer was used to determine the ventral surface topography of fixed metastatic mammary adenocarcinoma cells. The ventral surface of the cell was scanned by moving the sample with a piezoelectric stage. The height of the ventral surface, as well as the reflectivity, were determined using phase shifting interferometry. An overlay of a fluorescence image with the interference data shows that the prominent dark regions of the interference image correlate with the location of the paxillin.

Paper Details

Date Published: 23 February 2006
PDF: 7 pages
Proc. SPIE 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII, 60900F (23 February 2006); doi: 10.1117/12.647584
Show Author Affiliations
Michael Bambino, The Cooper Union for the Advancement of Science and Art (United States)
Ben Ovryn, Albert Einstein College of Medicine (United States)

Published in SPIE Proceedings Vol. 6090:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson, Editor(s)

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