Multiphoton high-resolution 3D imaging of Langerhans cells and keratinocytes in the mouse skin model adopted for epidermal powdered immunization

Langerhans cells (LCs) can be targeted with DNA-coated gold micro-projectiles ("Gene Gun") to induce potent cellular and humoral immune responses. It is likely that the relative volumetric distribution of LCs and keratinocytes within the epidermis impacts on the efficacy of Gene Gun immunization protocols. This study quantified the three-dimensional (3D) distribution of LCs and keratinocytes in the mouse skin model with a near-infrared multiphoton laser-scanning microscope (NIR-MPLSM). Stratum corneum (SC) and viable epidermal thickness measured with MPLSM was found in close agreement with conventional histology. LCs were located in the vertical plane at a mean depth of 14.9 mum, less than 3 mum above the dermo-epidermal boundary and with a normal histogram distribution. This likely corresponds to the fact that LCs reside in the suprabasal layer (stratum germinativum). The nuclear volume of keratinocytes was found to be approximately 1.4 times larger than that of resident LCs (88.6 mum3). Importantly, the ratio of LCs to keratinocytes in mouse ear skin (1:15) is more than three times higher than that reported for human breast skin (1:53). Accordingly, cross-presentation may be more significant in clinical Gene Gun applications than in pre-clinical mouse studies. These interspecies differences should be considered in pre-clinical trials using mouse models.

Bandwidth requirements for shallow, high-resolution seismic reflection

The optimum bandwidth for shallow, high-resolution seismic reflection differs from that required for conventional petroleum reflection. An understanding of this issue is essential for correct choice of acquisition instrumentation. Numerical modelling of simple Bowen Basin coal structures illustrates that, for high-resolution imaging, it is important to accurately record all frequencies up to the limit imposed by earth scattering. On the contrary, the seismic image is much less dependent on frequencies at the lower end of the spectrum. These quantitative observations support the use of specialised high-frequency geophones for high-resolution seismic imaging. Synthetic seismic inversion trials demonstrate that, irrespective of the bandwidth of the seismic data, additional low-frequency impedance control is essential for accurate inversion. Inversion provides no compelling argument for the use of conventional petroleum geophones in the high-resolution arena.