Abstract
It has recently been shown 1 that mutations in Filamin A and B genes produce a large spectrum of skeletal disorders in developing fetuses. However, high-resolution optical microscopy in cartilage growth plate using fluorescent antibody assays, which should elucidate molecular aspects of these disorders, is extremely difficult due to the high level of autofluoresce in this tissue. We apply multiphoton, confocal, lifetime and spectral microscopy to (i) image and characterize autofluorophores in chondrocytes and subtract their contributions to obtain a corrected antibody-marker fluorescence signal, and (ii) measure the interaction between Filamin A and B proteins by detecting the fluorescence resonance energy transfer (FRET) between markers of the two proteins. Taking advantage of the different fluorescence spectra of the endogenous and exogenous markers, we can significantly reduce the autofluorescence background. Preliminary results of the FRET experiments suggest no interaction between Filamin A and B proteins. However, developing of new antibodies targeting the carboxy-terminal immunoglobulin-like domain may be necessary to confirm this result. confocal microscopy, FRET, FLIM, autofluorescence, spectral, lifetime, cartilage, Filamin A and B.
Original language | English (US) |
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Title of host publication | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
Editors | D.V. Nicolau, J. Enderlein, R.C. Leif, D.L. Farkas, R. Raghavachari |
Pages | 75-81 |
Number of pages | 7 |
Volume | 5699 |
DOIs | |
State | Published - 2005 |
Event | Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III - San Jose, CA, United States Duration: Jan 24 2005 → Jan 27 2005 |
Other
Other | Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III |
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Country | United States |
City | San Jose, CA |
Period | 1/24/05 → 1/27/05 |
ASJC Scopus subject areas
- Engineering(all)