Second harmonic generation imaging via nonlinear endomicroscopy.

A compact endomicroscope is the only solution for transferring second harmonic generation (SHG) imaging into in vivo imaging and real time monitoring the content and structure of collagen. This is important for early diagnoses of different diseases associated with collagen change. A compact nonlinear endomicroscope using a double clad fiber (DCF) is newly employed in SHG imaging. The experiment shows the core of the DCF can maintain the linear polarization of the excitation laser beam in particular directions, and the degree of polarization of the excitation laser beam directly affects signal to noise ratio of SHG imaging. The nonlinear endomicroscope can display clear three dimensional (3D) SHG images of mouse tail tendon without the aid of contrast agents, which reveals the collagen fiber structure at different depths. The high resolution of SHG imaging from the endomicroscope shows that SHG imaging can reveal additional information about the orientation and degree of organisation of proteins and collagen fibers than two-photon-excited fluorescence imaging. Therefore SHG imaging offers endomicroscopy with additional channel of imaging for understanding more about biological phenomena.

Nonlinear endomicroscopy using a double-clad fiber coupler.

A double-clad fiber coupler is developed to be used in two-photon-excited fluorescence endomicroscopy to replace a dichroic mirror and separate the fluorescence signal from the excitation laser beam. With the double-clad fiber coupler, the endomicroscope becomes more compact, easier to be aligned, and more stable in alignment. The double-clad fiber coupler can transmit 62% of the excitation laser beam through the core. The fluorescence collection efficiency of the double-clad fiber coupler is 34%, which is, to the best of our knowledge, the highest fluorescence collection efficiency achieved by couplers used in two-photon-excited fluorescence endomicroscopes. As a result, the contrast of endomicroscopy imaging is enhanced.

A 0.4-mm-diameter probe for nonlinear optical imaging.

A miniaturized probe that possesses a diameter of 0.4 mm is developed for two-photon-excited fluorescence imaging. The miniaturized probe was manufactured by the collapse of air holes and the formation of a lens on the tip of a double-clad photonic crystal fiber (DCPCF) using electric arc discharging from a conventional fusion splicer. As a result, a femtosecond pulsed laser beam delivered by the DCPCF can be directly focused on a sample for two-photon fluorescence imaging. The numerical aperture of the lensed DCPCF is 0.12. The corresponding focal spot size is 6 microm, which is close to the diffraction limit. This 0.4-mm-diamter probe can provide clear two-photon-excited fluorescence images of 10-microm-diameter fluorescent microspheres.

Transmission simulation of coherent optical OFDM signals in WDM systems.

In this letter, we first present the theoretical basis for coherent optical OFDM systems in direct up/down conversion architecture. We then demonstrate the transmission performance through simulation for WDM systems with coherent optical OFDM (CO-OFDM) including the fiber nonlinearity effect. The results show that the system Q of the WDM channels at 10 Gb/s is over 13.0 dB for a transmission up to 4800 km of standard-single-mode-fiber (SSMF) without dispersion compensation. A novel technique of partial carrier filling (PCF) for improving the nonlinearity performance of the transmission is also presented. The system Q of the WDM channels with a filling factor of 50 % at 10 Gb/s is improved from 15.1 dB to 16.8 dB for a transmission up to 3200 km of SSMF without dispersion compensation.

Graphene oxide nanoparticles for enhanced photothermal cancer cell therapy under the irradiation of a femtosecond laser beam.

Nano-sized graphene and graphene oxide (GO) are promising for biomedical applications, such as drug delivery and photothermal therapy of cancer. It is observed in this work that the ultrafast reduction of GO nanoparticles (GONs) with a femtosecond laser beam creates extensive microbubbling. To understand the surface chemistry of GONs on the microbubble formation, the GONs were reduced to remove most of the oxygen-containing groups to get reduced GONs (rGONs). Microbubbling was not observed when the rGONs were irradiated by the laser. The instant collapse of the microbubbles may produce microcavitation effect that brings about localized mechanical damage. To understand the potential applications of this phenomenon, cancer cells labeled with GONs or rGONs were irradiated with the laser. Interestingly, the microbubbling effect greatly facilitated the destruction of cancer cells. When microbubbles were produced, the effective laser power was reduced to less than half of what is needed when microbubbling is absent. This finding will contribute to the safe application of femtosecond laser in the medical area by taking advantage of the ultrafast reduction of GONs. It may also find other important applications that need highly localized microcavitation effects.

Enhanced two-channel nonlinear imaging by a highly polarized supercontinuum light source generated from a nonlinear photonic crystal fiber with two zero-dispersion wavelengths.

Real-time monitoring the variation of chlorophyll distributions and cellular structures in leaves during plant growth provides important information for understanding the physiological statuses of plants. Two-photon-excited autofluorescence imaging and second harmonic generation imaging of leaves can be used for monitoring the nature intrinsic fluorophores distribution and cellular structures of leaves by the use of the near-infrared region of light which has minimal light absorption by endogenous molecules and thus increases tissue penetration. However, the two-photon absorption peak of intrinsic fluorophores of a ficus benjamina leaf is 50 nm away from the second harmonic generation excitation wavelength, which cannot be effectively excited by a femtosecond laser beam with one central wavelength. This paper shows that a highly polarized supercontinuum light generated from a birefringent nonlinear photonic crystal fiber with two zero-dispersion wavelengths can effectively excite two-photon autofluorescence as well as second harmonic generation signals for simultaneously monitoring intrinsic fluorophore distributions and non-centrosymmetric structures of leaves.

Reduction of self-phase modulation in double-clad photonic crystal fiber for nonlinear optical endoscopy.

Double-clad photonic crystal fiber and double-clad fiber have been widely used in multiphoton-excited fluorescence or second-harmonic generation (SHG) endoscopy. We provide a useful comparison of two fibers used in nonlinear optical microendoscopy. While a double-clad fiber is found to have a higher percentage of the output power from its core, which results in the efficient utilization of the power of the excitation laser, a double-clad photonic crystal fiber has a higher threshold of the nonlinearity, which effectively reduces the self-modulation effect and thus leads to a higher degree of polarization of the excitation beam. Consequently, the use of the double-clad photonic crystal fiber facilitates bright two-photon fluorescence imaging as well as polarized SHG imaging.

Imaging of goblet cells as a marker for intestinal metaplasia of the stomach by one-photon and two-photon fluorescence endomicroscopy.

Goblet cells are a requirement for the diagnosis of intestinal metaplasia of the stomach. The gastric mucosa is lined by a monolayer of columnar epithelium with some specialization at the crypts, but there are no goblet cells in normal gastric epithelium. The appearance of goblet cells in gastric epithelium is an indicator of potential malignant progression toward adenocarcinoma. Therefore, in vivo three-dimensional imaging of goblet cells is essential for diagnoses of a premalignant stage of gastric cancers called intestinal metaplasia. We used mouse intestine, which has goblet cells, as a model of intestinal metaplasia. One-photon confocal fluorescence endomicroscopy and two-photon fluorescence endomicroscopy are employed for 3-D imaging of goblet cells. The penetration depth, the sectioning ability, and the photobleaching information of imaging are demonstrated. Both endomicroscopy techniques can three-dimensionally observe goblet cells in mouse large intestine and achieve an imaging depth of 176 microm. The two-photon fluorescence endomicroscopy shows higher resolution and contrast of the imaging sections at each depth. In addition, two-photon fluorescence endomicroscopy also has advantages of sectioning ability and less photobleaching. These results prove that two-photon fluorescence endomicroscopy is advantageous in diagnoses of a premalignant stage of gastric cancers.

Fast handheld two-photon fluorescence microendoscope with a 475 microm x 475 microm field of view for in vivo imaging.

A fast handheld two-photon fiber-optic fluorescence endoscope for three-dimensional (3D) in vivo cellular imaging is developed. The compact handheld probe of the two-photon endoscope can simply be placed into contact with the target tissue to reveal clear 3D surface and subsurface histological images without biopsy. The new system has, to the best of our knowledge, the largest field of view (FOV) of 475 microm x 475 microm and a 3D imaging volume larger than 475 microm x 475 microm x 250 microm. A real-time two-photon fluorescence image is displayed at 0.4 mm(2)/s. The lateral and axial resolutions of the two-photon fluorescence endoscope are better than 1 and 14.5 microm, respectively.