Abstract

We report on a nonlinear optical microscope that adopts double-clad photonic crystal fibers for single-mode illumination delivery and multimode signal collection. It is demonstrated that two-photon fluorescence and second harmonic generation signals can be simultaneously collected in such a microscope with axial resolution of 2.8 µm and 2.5 µm, respectively. The delivery and detection efficiencies of the photonic-crystal-fiber-based microscope are significantly improved by approximately 3 times and 40 times compared with those in the single-mode fiber-optic microscope. The high resolution three-dimensional second harmonic generation images from rat tail tendon demonstrate the effectiveness of the system.

© 2005 Optical Society of America

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References

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Appl. Opt.

J. Microsc.

D. Bird and M. Gu, �??Fibre-optic two-photon scanning fluorescence microscopy,�?? J. Microsc. 208, 35-48 (2002).
[CrossRef] [PubMed]

Nat. Biotechnol.

W. R. Zipfel, R. M. Williams and W. W. Webb, �??Nonlinear magic: multiphoton microscopy in the biosciences,�?? Nat. Biotechnol. 21, 1369-1377 (2003).
[CrossRef] [PubMed]

P. J. Campagnola and L. M. Loew, �??Second harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,�?? Nat. Biotechnol. 21, 1356-1360 (2003).
[CrossRef] [PubMed]

Opt. Express

Opt. Lett.

Other

Bjarklev, J. Broeng, and A. S. Bjarklev, Photonic Crystal Fibers (Kluwer Academic Publishers, Norwell, Boston, 2003).
[CrossRef]

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Figures (5)

Fig. 1.
Fig. 1.

(a) Scanning electron microscopy image of a double-clad PCF. (b) Coupling efficiency of the double-clad PCF in the wavelength range 410–870 nm for three values of the NA of coupling objectives (0.07, 0.25 and 0.65). (c)–(h) Digital camera photograph of the output pattern from a double-clad PCF between 410 and 800 nm. A microscope objective with NA 0.07 is used for coupling. (i) Gaussian fit of an intensity profile at the output of the fiber. (j) Schematic diagram of the nonlinear optical microscope based on a double-clad PCF. ND, neutral density filter. Other abbreviations defined in the text.

Fig. 2.
Fig. 2.

Axial responses of the TPEF and SHG signals from a thin layer of AF-50 dye at an excitation wavelength of 800 nm. Inset shows the axial responses for TPEF in the case of the well-coupled illumination in the central core and the decoupled illumination in the inner cladding of the fiber. The power on the sample is approximately 1.5 mW.

Fig. 3.
Fig. 3.

Detected intensity of TPEF and SHG from the double-clad-PCF-based and a standard SMF-based microscope as a function of the power before the imaging objective.

Fig. 4.
Fig. 4.

SHG images from a scale of black tetra fish with (a) PCF-based microscope and (b) fiber-coupler-based microscope. Scale bars represent 20 µm.

Fig. 5.
Fig. 5.

(a)–(i) A series of SHG images sections from the rat tail tendon in the nonlinear optical microscope using a double-clad PCF. The image section spacing is 2 µm and the excitation power through the fiber core is approximately 5 mW. The scale bars in (a) represents 10 µm.

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