Values of integrated density and positive area in epidermis and dermis at 1 hour and 24 hour time points.

<p>* indicates p<0.05 and ** indicates p<0.01.</p

LSCM images of rigin delivery into excised human skin.

<p>Representative rigin (RIG) treated confocal laser scanning microscopy (CLSM) images of viable epidermis and dermis at 1 and 24 hour(s) are shown without and with microneedle (MN) delivery enhancement. Mosaic images of the viable epidermis and dermis (top row and bottom row, respectively) are shown at at 1 and 24 hour(s) after rigin delivery with microneedle enhancement. Each mosiac is 5×5 mm².</p

Integrated density and positive area data from melanostatin treated skin.

<p>Melanostatin (MEL) delivery characteristics are shown from epidermal (a and c) and dermal (d and d) mosiacs. Both the integrated density (a and b) and positive area (c and d) are shown for each microneedle site (MN). Data are shown for both 1 and 24 hours peptide exposure (1 h and 24 h, respecitvely). * indicates p<0.05. ** indicates p<0.01.</p

Integrated density and positive area data from rigin treated skin.

<p>Rigin (RIG) delivery characteristics are shown from epidermal (a and c) and dermal (d and d) mosiacs. Both the integrated density (a and b) and positive area (c and d) are shown for each microneedle site (MN). Data are shown for both 1 and 24 hours peptide exposure (1 h and 24 h, respecitvely). * indicates p<0.05. ** indicates p<0.01.</p

LSCM images of melanostatin delivery into excised human skin.

<p>Representative melanostatin (MEL) treated confocal laser scanning microscopy (CLSM) images of viable epidermis and dermis at 1 and 24 hour(s) are shown without and with microneedle (MN) delivery enhancement. Mosaic images of the viable epidermis and dermis (top row and bottom row, respectively) are shown at at 1 and 24 hour(s) after melanstatin delivery with microneedle enhancement. Each mosiac is 5×5 mm².</p

LSCM images of Pal-KTTKS delivery into excised human skin.

<p>Representative Pal-KTTKS treated confocal laser scanning microscopy (CLSM) images of viable epidermis and dermis at 1 and 24 hour(s) are shown without and with microneedle (MN) delivery enhancement. Mosaic images of the viable epidermis and dermis (top row and bottom row, respectively) are shown at at 1 and 24 hour(s) after Pal-KTTKS delivery with microneedle enhancement. Each mosiac is 5×5 mm².</p

THG power dependency plot.

<p>THG images of osteocytes obtained at 1550 nm (a) and 1700 nm (b) excitation <i>in vivo</i> as well as a logarithmic plot (c) of the THG intensity of an osteocyte with fundamental laser power for both 1550 nm and 1700 nm excitation. The data was fit with a line corresponding to a slope of 2.8 ± 0.2 for 1550 nm excitation and 2.9 ± 0.2 for 1700 nm excitation.</p

Images of a lacuna and canaliculi in the calvarial bone of a transgenic 2.3ColGFP mouse.

<p>THG (a) and 3PF of SR101 dye (b) were imaged with 1700 nm excitation while 2PF from GFP (c) was imaged with 850 nm excitation. A structural image cross-correlation analysis [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0186846#pone.0186846.ref031" target="_blank">31</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0186846#pone.0186846.ref032" target="_blank">32</a>] between 3PF, 2PF, and THG was performed (d).</p

A schematic of the laser microscope for three-photon imaging using 1550 nm and 1700 nm.

<p>Second and third harmonic generation (SHG and THG) as well as three photon excitation fluorescence (3PF) are collected while the laser is scanned with a polygonal and galvanometric mirror pair. HWP represents half-wave plates, PCF represents a polarization maintaining large mode area single mode photonic crystal fiber for generation of 1700 nm, while BiBO represents a bismuth borate crystal for frequency doubling of 1700 nm.</p

LCN parameters for WT and HDAC4/5 DKO mice.

<p>Measurement of the number of osteocytes in a volume consisting of 100 × 100 × 10 μm³ (a), lacunar surface area (b) and lacunar volume (d) per osteocyte as well as the number of canaliculi per lacunar surface area (e) in WT and HDAC4/5 DKO mice. Data are represented as the mean ± standard deviation (error bar). A statistically significant difference was not found in the lacunar surface area and volume measurements. The lacunar surface area, lacunar volume and number of canaliculi were determined from 3D renderings generated with Imaris software where an example osteocyte in WT (c) and HDAC4/5 DKO (f) mice is shown.</p