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Proceedings Paper

Biological applications of an LCoS-based programmable array microscope (PAM)
Author(s): Guy M. Hagen; Wouter Caarls; Martin Thomas; Andrew Hill; Keith A. Lidke; Bernd Rieger; Cornelia Fritsch; Bert van Geest; Thomas M. Jovin; Donna J. Arndt-Jovin
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Paper Abstract

We report on a new generation, commercial prototype of a programmable array optical sectioning fluorescence microscope (PAM) for rapid, light efficient 3D imaging of living specimens. The stand-alone module, including light source(s) and detector(s), features an innovative optical design and a ferroelectric liquid-crystal-on-silicon (LCoS) spatial light modulator (SLM) instead of the DMD used in the original PAM design. The LCoS PAM (developed in collaboration with Cairn Research, Ltd.) can be attached to a port of a(ny) unmodified fluorescence microscope. The prototype system currently operated at the Max Planck Institute incorporates a 6-position high-intensity LED illuminator, modulated laser and lamp light sources, and an Andor iXon emCCD camera. The module is mounted on an Olympus IX71 inverted microscope with 60-150X objectives with a Prior Scientific x,y, and z high resolution scanning stages. Further enhancements recently include: (i) point- and line-wise spectral resolution and (ii) lifetime imaging (FLIM) in the frequency domain. Multiphoton operation and other nonlinear techniques should be feasible. The capabilities of the PAM are illustrated by several examples demonstrating single molecule as well as lifetime imaging in live cells, and the unique capability to perform photoconversion with arbitrary patterns and high spatial resolution. Using quantum dot coupled ligands we show real-time binding and subsequent trafficking of individual ligand-growth factor receptor complexes on and in live cells with a temporal resolution and sensitivity exceeding those of conventional CLSM systems. The combined use of a blue laser and parallel LED or visible laser sources permits photoactivation and rapid kinetic analysis of cellular processes probed by photoswitchable visible fluorescent proteins such as DRONPA.

Paper Details

Date Published: 31 March 2007
PDF: 12 pages
Proc. SPIE 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, 64410S (31 March 2007); doi: 10.1117/12.710995
Show Author Affiliations
Guy M. Hagen, Max Planck Institute for Biophysical Chemistry (Germany)
Wouter Caarls, Delft Univ. of Technology (Netherlands)
Martin Thomas, Cairn Research Ltd. (United Kingdom)
Andrew Hill, Cairn Research Ltd. (United Kingdom)
Keith A. Lidke, Univ. of New Mexico (United States)
Bernd Rieger, Delft Univ. of Technology (Netherlands)
Cornelia Fritsch, Univ. of Sussex (United Kingdom)
Bert van Geest, Lambert Instruments (Netherlands)
Thomas M. Jovin, Max Planck Institute for Biophysical Chemistry (Germany)
Donna J. Arndt-Jovin, Max Planck Institute for Biophysical Chemistry (Germany)


Published in SPIE Proceedings Vol. 6441:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V
Daniel L. Farkas; Robert C. Leif; Dan V. Nicolau, Editor(s)

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