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 | |
| 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 |
Fingerprint
Keywords
- Autofluorescence
- Cartilage
- Confocal microscopy
- Filamin A and B
- Two-photon imaging
ASJC Scopus subject areas
- Electrical and Electronic Engineering
- Condensed Matter Physics
Cite this
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
}
TY - GEN
T1 - Confocal and two-photon imaging in cartilage
T2 - Expression patterns of Filamin A and B
AU - Wachsmann-Hogiu, Sebastian
AU - Krakow, Deborah
AU - Sebald, Eiman T.
AU - Bertolotto, Cristina
AU - Acuna, Dora
AU - Farkas, Daniel L.
PY - 2004
Y1 - 2004
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.
KW - Autofluorescence
KW - Cartilage
KW - Confocal microscopy
KW - Filamin A and B
KW - Two-photon imaging
UR - http://www.scopus.com/inward/record.url?scp=5644239742&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=5644239742&partnerID=8YFLogxK
U2 - 10.1117/12.561883
DO - 10.1117/12.561883
M3 - Conference contribution
VL - 5322
SP - 140
EP - 145
BT - Proceedings of SPIE - The International Society for Optical Engineering
A2 - Nicolau, D.V.
A2 - Enderlein, J.
A2 - Leif, R.C.
A2 - Farkas, D.L.
ER -