Measuring the surface topography at focal adhesions using laser feedback interferometry

Michael Bambino; Ben Ovryn

doi:10.1117/12.647584

23 February 2006 Measuring the surface topography at focal adhesions using laser feedback interferometry

Michael Bambino, Ben Ovryn

Proceedings Volume 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII; 60900F (2006) https://doi.org/10.1117/12.647584
Event: SPIE BiOS, 2006, San Jose, California, United States

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.

Citation Download Citation

Michael Bambino and Ben Ovryn "Measuring the surface topography at focal adhesions using laser feedback interferometry", Proc. SPIE 6090, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII, 60900F (23 February 2006); https://doi.org/10.1117/12.647584

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KEYWORDS

Reflectivity

Microscopes

Visibility

Geometrical optics

Luminescence

Microscopy

Interfaces

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