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

doi:10.1117/12.561883

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

Pathology and Laboratory Medicine

Research output: Chapter in Book/Report/Conference proceeding › Conference 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 language	English (US)
Title of host publication	Proceedings of SPIE - The International Society for Optical Engineering
Editors	D.V. Nicolau, J. Enderlein, R.C. Leif, D.L. Farkas
Pages	140-145
Number of pages	6
Volume	5322
DOIs	https://doi.org/10.1117/12.561883
State	Published - 2004
Event	Progress in Biomedical Optics and Imaging - Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II - San Jose, CA, United States Duration: Jan 27 2004 → Jan 28 2004

Other

Other	Progress in Biomedical Optics and Imaging - Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II
Country	United States
City	San Jose, CA
Period	1/27/04 → 1/28/04

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

Confocal and two-photon imaging in cartilage : Expression patterns of Filamin A and B. / Wachsmann-Hogiu, Sebastian; Krakow, Deborah; Sebald, Eiman T.; Bertolotto, Cristina; Acuna, Dora; Farkas, Daniel L.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / D.V. Nicolau; J. Enderlein; R.C. Leif; D.L. Farkas. Vol. 5322 2004. p. 140-145.

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Wachsmann-Hogiu, S, Krakow, D, Sebald, ET, Bertolotto, C, Acuna, D & Farkas, DL 2004, Confocal and two-photon imaging in cartilage: Expression patterns of Filamin A and B. in DV Nicolau, J Enderlein, RC Leif & DL Farkas (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 5322, pp. 140-145, Progress in Biomedical Optics and Imaging - Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II, San Jose, CA, United States, 1/27/04. https://doi.org/10.1117/12.561883

Wachsmann-Hogiu, Sebastian ; Krakow, Deborah ; Sebald, Eiman T. ; Bertolotto, Cristina ; Acuna, Dora ; Farkas, Daniel L. / Confocal and two-photon imaging in cartilage : Expression patterns of Filamin A and B. Proceedings of SPIE - The International Society for Optical Engineering. editor / D.V. Nicolau ; J. Enderlein ; R.C. Leif ; D.L. Farkas. Vol. 5322 2004. pp. 140-145

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N2 - 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.

AB - 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.

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