Biological applications of an LCoS-based programmable array microscope (PAM)

Guy M. Hagen; Wouter Caarls; Martin Thomas; Andrew Hill; Keith A. Lidke; Bernd Rieger; Cornelia Fritsch; Bert van Geest; Thomas M. Jovin M.D.; Donna J. Arndt-Jovin

doi:10.1117/12.710995

31 March 2007 Biological applications of an LCoS-based programmable array microscope (PAM)

Guy M. Hagen, Wouter Caarls, Martin Thomas, Andrew Hill, Keith A. Lidke, Bernd Rieger, Cornelia Fritsch, Bert van Geest, Thomas M. Jovin M.D., Donna J. Arndt-Jovin

Author Affiliations +

Proceedings Volume 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V; 64410S (2007) https://doi.org/10.1117/12.710995
Event: SPIE BiOS, 2007, San Jose, California, United States

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.

Citation Download Citation

Guy M. Hagen, Wouter Caarls, Martin Thomas, Andrew Hill, Keith A. Lidke, Bernd Rieger, Cornelia Fritsch, Bert van Geest, Thomas M. Jovin M.D., and Donna J. Arndt-Jovin "Biological applications of an LCoS-based programmable array microscope (PAM)", Proc. SPIE 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, 64410S (31 March 2007); https://doi.org/10.1117/12.710995

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available