0

Full Content is available to subscribers

Subscribe/Learn More  >
Proceedings Article

Fiber-coupled non-descanned 4π detection with a commercial confocal microscope modified for multiphoton imaging

[+] Author Affiliations
Adam M. Larson

Univ. of Rochester (USA)

Vijay Iyer

Rice Univ. (USA)

Tycho Hoogland

Baylor College of Medicine (USA)

Peter Saggau

Baylor College of Medicine (USA) and Rice Univ. (USA)

Proc. SPIE 4963, Multiphoton Microscopy in the Biomedical Sciences III, 239 (July 9, 2003); doi:10.1117/12.478000
Text Size: A A A
From Conference Volume 4963

  • Multiphoton Microscopy in the Biomedical Sciences III
  • Ammasi Periasamy; Peter T. C. So
  • San Jose, CA | January 25, 2003

abstract

The extremely small (femtoliter) excitation volume of multiphoton (MP) microscopy renders all emitted photons useful in detecting fluorescence signals. Hence, multiphoton laser scanning microscopy (MPLSM) systems can collect fluorescence through the objective (epi-fluorescence), as well as the condenser (trans-fluorescence). For maximal collection efficiency, both optical paths can be used concurrently (4pi detection). Most MPLSM systems incorporate photodetectors directly in or adjacent to the epi- and trans-fluorescence optical paths of the microscope, generally photomultiplier tubes with associated optics. These arrangements are optically straightforward, but are often bulky and difficult to reconfigure. Here, we demonstrate that all fluorescence from the specimen can be efficiently coupled into two multimode optical fibers-one each for the epi- and trans-fluorescence pathways. Fiber-coupled detection enables a modular detection paradigm where light can be routed to easily reconfigurable and interchangeable detection module(s). A novel MPLSM system was constructed, which is readily switched between the original de-scanned detection path for confocal microscopy, and the newly added pathways supporting fiber-coupled non-descanned 4pi detection for MP microscopy. Sample MP images of fluorescent beads and fluorescent-labeled hippocampal neurons are presented, demonstrating the viability of fiber-coupled detection.

© (2003) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
Citation

Adam M. Larson ; Vijay Iyer ; Tycho Hoogland and Peter Saggau
"Fiber-coupled non-descanned 4π detection with a commercial confocal microscope modified for multiphoton imaging", Proc. SPIE 4963, Multiphoton Microscopy in the Biomedical Sciences III, 239 (July 9, 2003); doi:10.1117/12.478000; http://dx.doi.org/10.1117/12.478000


Access This Article
Sign In to Access Full Content
Please Wait... Processing your request... Please Wait.
Sign in or Create a personal account to Buy this article ($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


Buy this article ($18 for members, $25 for non-members).
Sign In