Abstract
The W. M. Keck 3-Dimensional Fusion Microscope's (3DFM) multimodal imaging platform provides researchers the unique ability to capture and process images from 5 co-registered optical imaging modalities without moving the specimen. Here, we detail the addition and first experimental results of a novel 6th modality, multiphoton fluorescence anisotropy, which allows for dynamic studies of macromolecule binding interactions. We have begun the application of Fluorescence Lifetime Intensity Microscopy (FLIM) techniques using a Time Correlated Single Photon Counting (TCSPC) module to measure the timedependent decay of polarized fluorescence. Calculation of the time-dependent fluorescence anisotropy decay provides information related to the rotational dynamics of the excited fluorophores. Data corresponding to the calibration of the excitation and emission polarization planes, sample images of fluorescence anisotropy measured from a randomly labeled sample, and initial results of fluorescence anisotropy decay experiments are presented.
Keywords
Polarimetric Multiphoton Imaging, Fusion Microscope, anisotropy, 3DFM, TCSPC
Subject Categories
Three-dimensional imaging, Anisotropy
Disciplines
Biochemistry, Biophysics, and Structural Biology
Publisher
Bernard M. Gordon Center for Subsurface Sensing and Imaging Systems (Gordon-CenSSIS)
Publication Date
2007
Rights Holder
Bernard M. Gordon Center for Subsurface Sensing and Imaging Systems (Gordon-CenSSIS)
Permanent URL
Recommended Citation
Bouchard, Matthew B.; Warger II, William C.; Laevksy, Gary S.; and DiMarzio, Charles A., "Adding polarimetric multiphoton imaging to the W. M. Keck 3-dimensional fusion microscope" (2007). BioBED Presentations. Paper 1. http://hdl.handle.net/2047/d1000919x
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COinS
Notes
Poster presented at the 2007 Validating TestBED and Research on Real World Problems for the I-PLUS Development Conference.