Full Content is available to subscribers

Subscribe/Learn More  >
Proceedings Article

Microspectroscopic method for determination of size and distribution of protein complexes in vivo

[+] Author Affiliations
S. Rath, A. P. Sullivan, M. R. Stoneman, V. Raicu

Univ. of Wisconsin-Milwaukee (USA)

Proc. SPIE 7378, Scanning Microscopy 2009, 737829 (May 22, 2009); doi:10.1117/12.821831
Text Size: A A A
From Conference Volume 7378

  • Scanning Microscopy 2009
  • Michael T. Postek; Dale E. Newbury; S. Frank Platek; David C. Joy
  • Monterey, CA | May 04, 2009

abstract

Resonant Energy Transfer (RET) from an optically excited molecule to a non-excited molecule residing nearby has been used to detect molecular interactions in living cells. Information such as the number of proteins forming a molecular complex has been obtained so far for a handful of proteins, but only after exposing the samples sequentially to at least two different excitation wavelengths. Changes in the molecular makeup of a cellular region occurring during this lengthy process of measurement has limited the applicability of RET to determination of cellular averages. We developed a method for imaging protein complex distribution in living cells with sub-cellular spatial resolution, which relies on a spectrally-resolved two-photon microscope. The use of diffractive optics in a non-descanned configuration allows acquisition of a full set of spectrally-resolved images after only one complete scan of the excitation beam. This presentation will briefly describe our basic experimental setup and a simple theory of RET in oligomeric complexes, and it will review our recent results on determination of the geometry and size of oligomeric complexes of several proteins in yeast as well as in mammalian cells. This method basically transforms RET into a method for performing veritable structural determinations of protein complexes in vivo.

© (2009) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
Citation

S. Rath ; A. P. Sullivan ; M. R. Stoneman and V. Raicu
"Microspectroscopic method for determination of size and distribution of protein complexes in vivo", Proc. SPIE 7378, Scanning Microscopy 2009, 737829 (May 22, 2009); doi:10.1117/12.821831; http://dx.doi.org/10.1117/12.821831


Access This Proceeding
Sign in or Create a personal account to Buy this proceeding ($15 for members, $18 for non-members).

Figures

Tables

NOTE:
Citing articles are presented as examples only. In non-demo SCM6 implementation, integration with CrossRef’s "Cited By" API will populate this tab (http://www.crossref.org/citedby.html).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Proceeding
Sign in or Create a personal account to Buy this proceeding ($15 for members, $18 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.