Multiphoton, confocal, and lifetime microscopy for molecular imaging in cartilage

Sebastian Wachsmann-Hogiu, Deborah Krakow, Veneta Kirilova, Daniel H. Cohn, Cristina Bertolotto, Dora Acuna, Qiyin Fang, Nikola Krivorov, Daniel L. Farkas

Research output: Chapter in Book/Report/Conference proceedingConference contribution

3 Scopus citations

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 languageEnglish (US)
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
EditorsD.V. Nicolau, J. Enderlein, R.C. Leif, D.L. Farkas, R. Raghavachari
Pages75-81
Number of pages7
Volume5699
DOIs
StatePublished - 2005
EventImaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III - San Jose, CA, United States
Duration: Jan 24 2005Jan 27 2005

Other

OtherImaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III
CountryUnited States
CitySan Jose, CA
Period1/24/051/27/05

ASJC Scopus subject areas

  • Engineering(all)

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    Wachsmann-Hogiu, S., Krakow, D., Kirilova, V., Cohn, D. H., Bertolotto, C., Acuna, D., Fang, Q., Krivorov, N., & Farkas, D. L. (2005). Multiphoton, confocal, and lifetime microscopy for molecular imaging in cartilage. In D. V. Nicolau, J. Enderlein, R. C. Leif, D. L. Farkas, & R. Raghavachari (Eds.), Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 5699, pp. 75-81). [11] https://doi.org/10.1117/12.597670