, Tokyo,

Japan). The MN arrays were also visualised using a Phoenix X-ray nanotom system (GE, London, UK), under the inhibitors following conditions; energy: 55 kV, current: 160 mA, nanotom mode: 0, voxel resolution: 10 μm, number of projections: 720, image averaging: 3, detector timing: 1500 ms, binning mode: 1 × 1 (no binning). The method involved the acquisition of a series of X-ray projection images at a known number of angular positions through 360°. Variation in the contrast of each projection image relates to how the x-rays are attenuated as they penetrate the sample. Axial slice views were computed from the X-ray projections using back projection reconstruction algorithms. 3D rendering of the all axial slice views allowed visualisation of VRT752271 in vivo the 3D model. He-ion images of the MN arrays were generated using the Orion Helium- ion microscope (Carl Zeiss Smt GmBH, Oberkochen, Germany). He-ion technology relies on a novel high brightness helium ion source of atomic dimensions. The helium beam was focused on the sample with an ultimate probe size of 0.25 nm. The images provide rich surface–specific

information due to the unique nature of the beam-sample interaction. The hollow MNs were imaged under selleck products the following conditions: Acceleration 29.0 kV, dwell time 1.0 μs, blanker current 6.7 pA, working distance 22–23 mm. The force required to successfully insert the PC MNs into excised porcine skin was determined using a TA.XT-plus Texture Analyser (Stable Micro Systems, Surrey, UK). Neonate porcine skin was obtained from stillborn piglets and immediately (<24 h after those birth) excised and trimmed to a thickness of approximately 400 μm using an electric dermatome (Integra Life Sciences™, Padgett Instruments, NJ, USA). Skin was then stored in aluminium foil

at −20 °C until further use but for no longer than 2 weeks. Skin was equilibrated in PBS for an hour and hair was removed using a disposable razor. The SC surface of the skin was dried with tissue paper and the skin was placed, dermis side down, on a 500 μm-thick sheet of dental wax, and this assembly was then secured on a wooden block for support. MNs were attached to the tip of a moveable cylindrical probe (length 5 cm, cross-sectional area 1.5 cm2) using cyanoacrylate adhesive (Loctite, Dublin, Ireland). The test station, in compression mode, then pressed MN arrays against the skin at a speed of 0.5 mm/s for 30 s with known forces of 0.05, 0.10 and 0.40 N/needle. Following MN removal, methylene blue solution (1% w/v) was applied onto the skin surface and left for 15 min. This solution was then gently wiped off, first with dry tissue paper and then with saline and alcohol swabs. The surface of the stained skin was then photographed using a digital camera (Nikon Coolpix I120®, Nikon UK Ltd., Surrey, UK) and the number of methylene blue stained micro-conduits was simply counted.