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Proceedings Paper

Multiphoton, confocal, and lifetime microscopy for molecular imaging in cartilage
Author(s): Sebastian Wachsmann-Hogiu; Deborah Krakow; Veneta T. Kirilova; Daniel H. Cohn; Cristina Bertolotto; Dora Acuna; Qiyin Fang; Nikola Krivorov; Daniel L. Farkas
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Paper Abstract

It has recently been shown 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.

Paper Details

Date Published: 29 March 2005
PDF: 7 pages
Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005); doi: 10.1117/12.597670
Show Author Affiliations
Sebastian Wachsmann-Hogiu, Cedars-Sinai Medical Ctr. (United States)
Deborah Krakow, Cedars-Sinai Medical Ctr. (United States)
David Geffen School of Medicine at UCLA (United States)
Veneta T. Kirilova, Cedars-Sinai Medical Ctr. (United States)
Daniel H. Cohn, Cedars-Sinai Medical Ctr. (United States)
David Geffen School of Medicine at UCLA (United States)
Cristina Bertolotto, Cedars-Sinai Medical Ctr. (United States)
David Geffen School of Medicine at UCLA (United States)
Dora Acuna, Cedars-Sinai Medical Ctr. (United States)
Qiyin Fang, Cedars-Sinai Medical Ctr. (United States)
Nikola Krivorov, Cedars-Sinai Medical Ctr. (United States)
Daniel L. Farkas, Cedars-Sinai Medical Ctr. (United States)


Published in SPIE Proceedings Vol. 5699:
Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III
Dan V. Nicolau; Dan V. Nicolau; Jörg Enderlein; Ramesh Raghavachari; Robert C. Leif; Daniel L. Farkas, Editor(s)

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