Imaging in thick samples, a phased Monte Carlo radiation transfer algorithm.

SIGNIFICANCE: Optical microscopy is characterized by the ability to get high resolution, below 1 µm, high contrast, functional and quantitative images. The use of shaped illumination, such as with lightsheet microscopy, has led to greater three-dimensional isotropic resolution with low phototoxicity. However, in most complex samples and tissues, optical imaging is limited by scattering. Many solutions to this issue have been proposed, from using passive approaches such as Bessel beam illumination to active methods incorporating aberration correction, but making fair comparisons between different approaches has proven to be challenging. AIM: We present a phase-encoded Monte Carlo radiation transfer algorithm (φMC) capable of comparing the merits of different illumination strategies or predicting the performance of an individual approach. APPROACH: We show that φMC is capable of modeling interference phenomena such as Gaussian or Bessel beams and compare the model with experiment. RESULTS: Using this verified model, we show that, for a sample with homogeneously distributed scatterers, there is no inherent advantage to illuminating a sample with a conical wave (Bessel beam) instead of a spherical wave (Gaussian beam), except for maintaining a greater depth of focus. CONCLUSION: φMC is adaptable to any illumination geometry, sample property, or beam type (such as fractal or layered scatterer distribution) and as such provides a powerful predictive tool for optical imaging in thick samples.

The application of optical coherence tomography to image subsurface tissue structure of Antarctic krill Euphausia superba.

Many small open ocean animals, such as Antarctic krill, are an important part of marine ecosystems. To discover what will happen to animals such as krill in a changing ocean, experiments are run in aquaria where conditions can be controlled to simulate water characteristics predicted to occur in the future. The response of individual animals to changing water conditions can be hard to observe, and with current observation techniques it is very difficult to follow the progress of an individual animal through its life. Optical coherence tomography (OCT) is an optical imaging technique that allows images at high resolution to be obtained from depths up to a few millimeters inside biological specimens. It is compatible with in vivo imaging and can be used repeatedly on the same specimens. In this work, we show how OCT may be applied to post mortem krill samples and how important physiological data such as shell thickness and estimates of organ volume can be obtained. Using OCT we find an average value for the thickness of krill exoskeleton to be (30±4) µm along a 1 cm length of the animal body. We also show that the technique may be used to provide detailed imagery of the internal structure of a pleopod joint and provide an estimate for the heart volume of (0.73±0.03) mm3.

Fundamentally mode-locked, femtosecond waveguide oscillators with multi-gigahertz repetition frequencies up to 15 GHz.

We demonstrate passively mode-locked Yb(3+)-doped glass waveguide lasers in a quasi-monolithic configuration with a maximum pulse repetition frequency up to 15.2 GHz. A semiconductor saturable absorber mirror (SESAM) is used to achieve stable mode-locking around 1050 nm with pulse durations as short as 811 fs and an average power up to 27 mW. Different waveguide samples are also employed to deliver pulses with repetition rates of 4.9 GHz, 10.4 GHz and 12 GHz with an average power of 32 mW, 60 mW and 45 mW, respectively. The group velocity dispersion control in the cavity is provided by changing the gap between the SESAM and the waveguide end-face to facilitate a soliton mode-locking regime.

Discrimination of normal from pre-malignant cervical tissue by Raman mapping of de-paraffinized histological tissue sections.

The authors present Raman cluster mapping of de-paraffinized normal cervical tissue and demonstrate the ability of this approach to differentiate between normal squamous epithelium and cervical intraepithelial neoplasia (CIN). Multivariate analysis was performed by hierarchical cluster analysis (HCA) of the Raman spectra associated with the different tissue types and Raman maps were generated using the resultant clusters. Using normal cervical tissue, squamous epithelium and the epithelial-stromal interface, a muscular artery and endocervical glands were successfully mapped. Analysis of a tissue section containing a cervical intraepithelial neoplasia (CIN) grade 2 lesion adjacent to normal squamous epithelium demonstrated that the CIN lesion clustered predominantly with the basal epithelial cells of normal epithelium and allowed visual discrimination of these areas using the Raman map. These findings suggest that Raman mapping has the potential to provide images that are useful for disease diagnosis. In particular, the discrimination between normal cervical squamous epithelium and CIN is of relevance to cervical screening pathology.

Targeted optical injection of gold nanoparticles into single mammalian cells.

We present an all optical technique for the targeted delivery of single 100 nm diameter gold nanoparticles into a specified region of the interior of an individual mammalian cell through a combination of optical tweezing and optical injection. The internalisation of the nanoparticle is verified by confocal laser scanning microscopy and confocal laser scanning reflectance microscopy. This represents the first time that nano sized particles have been tweezed and optically injected into mammalian cells using only light, and provides a novel methodology for internalising nanosphere based biosensors within specific intracellular regions of a mammalian cell.

Enhanced operation of femtosecond lasers and applications in cell transfection.

In this work we present a review and discussion on the enhancement of femtosecond (fs) lasers for use within biophotonics with a particular focus on their use in optical transfection techniques. We describe the broad range of source options now available for the generation of femtosecond pulses before briefly reviewing the application of fs laser in optical transfection studies. We show that major performance enhancements may be obtained by optimising the spatial and temporal performance of the laser source before considering possible future directions in this field. In relation to optical transfection we describe how such laser sources initiate a multiphoton process to permeate the cell membrane in a transient fashion. We look at aspects of this technique including the ability to combine transfection with optical trapping. For future implementation of such transfection we explore the role of new sources and "nondiffracting" light fields.