Confocal and two-photon imaging in cartilage: Expression patterns of Filamin A and B

Sebastian Wachsmann-Hogiu, Deborah Krakow, Eiman T. Sebald, Cristina Bertolotto, Dora Acuna, Daniel L. Farkas

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

1 Scopus citations

Abstract

Optical imaging in cartilage is challenging due to the high levels of intra- and inter-cellular autofluorescence. We report here on high-resolution confocal and two-photon imaging of endogenous fluorescence of cartilage and of exogenous fluorescence of filamin A and B protein markers. Confocal laser scanning microscopy offers the advantage of quasi-theoretical spatial resolution and minimizes the autofluorescence contribution by eliminating the out-of-focus light. In non-labeled cartilage, we observe mostly intracellular autofluorescence that, due to the uniform distribution within the cell, can be further effectively minimized by careful choice of experimental parameters. The fluorescence of the exogenous markers AlexaFluor 488 and AlexaFluor 568 labeling Filamins A and B, respectively, could also be detected and quantitated using this procedure, revealing topologically different expression levels of filamin A and B proteins in the cartilage growth plate. Two-photon excited fluorescence imaging yielded further resolution improvements and structural and functional information.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsD.V. Nicolau, J. Enderlein, R.C. Leif, D.L. Farkas
Pages140-145
Number of pages6
Volume5322
DOIs
StatePublished - 2004
EventProgress in Biomedical Optics and Imaging - Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II - San Jose, CA, United States
Duration: Jan 27 2004Jan 28 2004

Other

OtherProgress in Biomedical Optics and Imaging - Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II
CountryUnited States
CitySan Jose, CA
Period1/27/041/28/04

Keywords

  • Autofluorescence
  • Cartilage
  • Confocal microscopy
  • Filamin A and B
  • Two-photon imaging

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

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    Wachsmann-Hogiu, S., Krakow, D., Sebald, E. T., Bertolotto, C., Acuna, D., & Farkas, D. L. (2004). Confocal and two-photon imaging in cartilage: Expression patterns of Filamin A and B. In D. V. Nicolau, J. Enderlein, R. C. Leif, & D. L. Farkas (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 5322, pp. 140-145) https://doi.org/10.1117/12.561883