Feasibility of tracking phospholipid permeation into skin using infrared and Raman microscopic imaging

The feasibility of monitoring the permeation of chain perdeuterated 1,2-dipalmitoylphosphatidylcholine (DPPC-d62) and 1-palmitoyl-d31, 2-oleoylphosphatidylcholine (P-d31OPC) vesicles into pigskin using infrared (IR) microscopic imaging and confocal Raman microscopy was demonstrated. The former technique permits the examination of the relative concentration of molecular species (e.g., endogenous and exogenous lipids and proteins) over spatial areas, approximately 1 mm, with a spatial resolution of approximately 10-12 microm. In contrast, Raman microscopy allows the confocal examination of tissue at depths up to 100 microm with a pixel size of about 2-3 microm3. Spectral signal/noise, however, is reduced from IR and significantly smaller areas are generally monitored. The permeation of the gel phase DPPC-d62 was limited to approximately 5-15 microm, whereas the liquid-crystalline phase P-d31OPC permeated to substantially greater depths (35-100 microm), at times ranging up to 24 h after application. The results are generally in accord with literature values. In addition, the state of the P-d31OPC (intact vesicles or molecularly dispersed with skin constituents) was evaluated from the spatial dependence of the deuteriopalmitate chain conformational order. Upon permeation, the chains became more ordered. The advantages and limitations of these imaging technologies are discussed.