Extraction of near-field fluorescence from composite signals to provide high resolution images of glial cells

Biophysical Journal
R T DoyleP G Haydon

Abstract

The subdiffraction optical resolution that can be achieved using near-field optical microscopy has the potential to permit new approaches and insights into subcellular function and molecular dynamics. Despite the potential of this technology, it has been difficult to apply to cellular samples. One significant problem is that sample thickness causes the optical information to be comprised of a composite signal containing both near- and far-field fluorescence. To overcome this issue we have developed an approach in which a near-field optical fiber is translated toward the cell surface. The increase in fluorescence intensity during z-translation contains two components: a far-field fluorescence signal when the tip of the fiber is distant from the labeled cell, and combined near- and far-field fluorescence when the tip interacts with the cell surface. By fitting a regression curve to the far-field fluorescence intensity as the illumination aperture approaches the cell, it is possible to isolate near-field from far-field fluorescent signals. We demonstrate the ability to resolve actin filaments in chemically fixed, hydrated glial cells. A comparison of composite fluorescence signals with extracted near-field fluorescence demonstrate...Continue Reading

References

Jun 30, 1994·Nature·V ParpuraP G Haydon
May 30, 1997·Annals of the New York Academy of Sciences·S P Marchese-Ragona, P G Haydon
Mar 24, 1999·Trends in Cell Biology·A LewisK Lieberman
Mar 30, 1999·Biophysical Journal·S A Vickery, R C Dunn
Sep 7, 1999·Journal of Neuroscience Methods·J D BuiW Tan
Jun 30, 1972·Nature·E A Ash, G Nicholls
Nov 26, 1993·Science·E Betzig, R J Chichester
Jul 15, 1994·Science·X S Xie, R C Dunn

❮ Previous
Next ❯

Citations

Mar 12, 2008·Journal of Microscopy·M ZweyerS Prato
Dec 10, 2003·Applied Optics·Alexandra FragolaClaude Boccara
Dec 13, 2005·Experimental Neurology·Etsuro Uemura, Heather West Greenlee

❮ Previous
Next ❯

Related Concepts

Related Feeds

Astrocytes

Astrocytes are glial cells that support the blood-brain barrier, facilitate neurotransmission, provide nutrients to neurons, and help repair damaged nervous tissues. Here is the latest research.