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Two-color, two-photon, and excited-state absorption microscopy.

Publication ,  Journal Article
Fu, D; Ye, T; Matthews, TE; Yurtsever, G; Warren, WS
Published in: Journal of biomedical optics
September 2007

We develop a new approach in imaging nonfluorescent species with two-color two-photon and excited state absorption microscopy. If one of two synchronized mode-locked pulse trains at different colors is intensity modulated, the modulation transfers to the other pulse train when nonlinear absorption takes places in the medium. We can easily measure 10(-6) absorption changes caused by either two-photon absorption or excited-state absorption with a RF lock-in amplifier. Sepia melanin is studied in detail as a model system. Spectroscopy studies on the instantaneous two-photon absorption (TPA) and the relatively long-lived excited-state absorption (ESA) of melanin are carried out in solution, and imaging capability is demonstrated in B16 cells. It is found that sepia melanin exhibits two distinct excited states with different lifetimes (one at 3 ps, one lasting hundreds of nanoseconds) when pumped at 775 nm. Its characteristic TPA/ESA enables us to image its distribution in cell samples with high resolution comparable to two-photon fluorescence microscopy (TPFM). This new technique could potentially provide valuable information in diagnosing melanoma.

Duke Scholars

Published In

Journal of biomedical optics

DOI

EISSN

1560-2281

ISSN

1083-3668

Publication Date

September 2007

Volume

12

Issue

5

Start / End Page

054004

Related Subject Headings

  • Sensitivity and Specificity
  • Reproducibility of Results
  • Optics
  • Microscopy, Fluorescence, Multiphoton
  • Microscopy, Confocal
  • Mice
  • Melanoma
  • Melanins
  • Image Enhancement
  • Equipment Failure Analysis
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Fu, D., Ye, T., Matthews, T. E., Yurtsever, G., & Warren, W. S. (2007). Two-color, two-photon, and excited-state absorption microscopy. Journal of Biomedical Optics, 12(5), 054004. https://doi.org/10.1117/1.2780173
Fu, Dan, Tong Ye, Thomas E. Matthews, Gunay Yurtsever, and Warren S. Warren. “Two-color, two-photon, and excited-state absorption microscopy.Journal of Biomedical Optics 12, no. 5 (September 2007): 054004. https://doi.org/10.1117/1.2780173.
Fu D, Ye T, Matthews TE, Yurtsever G, Warren WS. Two-color, two-photon, and excited-state absorption microscopy. Journal of biomedical optics. 2007 Sep;12(5):054004.
Fu, Dan, et al. “Two-color, two-photon, and excited-state absorption microscopy.Journal of Biomedical Optics, vol. 12, no. 5, Sept. 2007, p. 054004. Epmc, doi:10.1117/1.2780173.
Fu D, Ye T, Matthews TE, Yurtsever G, Warren WS. Two-color, two-photon, and excited-state absorption microscopy. Journal of biomedical optics. 2007 Sep;12(5):054004.

Published In

Journal of biomedical optics

DOI

EISSN

1560-2281

ISSN

1083-3668

Publication Date

September 2007

Volume

12

Issue

5

Start / End Page

054004

Related Subject Headings

  • Sensitivity and Specificity
  • Reproducibility of Results
  • Optics
  • Microscopy, Fluorescence, Multiphoton
  • Microscopy, Confocal
  • Mice
  • Melanoma
  • Melanins
  • Image Enhancement
  • Equipment Failure Analysis