Abstract
Confocal laser scanning microscopy (CLSM) is a well-used microscopic tool that provides valuable morphological and functional information within cells and tissues. The application of CLSM to skin and the topical penetration of nanoparticles (NP) will be addressed. First, we describe the advantages of confocal microscopy compared to other techniques and its use relative to skin research. Second, we discuss the ability of CLSM to detect single NP. Regarding their interaction with skin, the appropriate method to retain nanoparticle localization in the tissue with minimal fixation is critically important. Also, the interaction of several different types of NP (quantum dots, fullerene and dendrimers) and their interaction with skin detected by CLSM under various conditions (flexed, tape stripped and abraded skin) is reviewed. Finally, human epidermal keratinocytes and dendritic cells that serve as appropriate in vitro models for skin cell interactions and cellular uptake of NP are also discussed. In conclusion, the unique functions of CLSM such as the ability to detect fluorescence, optical sectioning, three dimensional remodeling, as well as its use in the reflection mode in tandem with other methods, provides great promise with broad applications regarding the interactions of nanomaterials with skin.
Highlights
Confocal laser scanning microscopy (CLSM) is a well-used microscopic tool that provides valuable morphological and functional information within cells and tissues
Polystyrene NP were applied on the skin of the porcine ear and found to accumulate preferentially in the follicular openings.[8]. Both studies included a phosphate buffered saline washing step following cryo-sectioning which may have altered the NP localization in the skin. Methods such as electron microscopy and energy-dispersive x-ray (EDX) used to study NP within skin may create more artifacts due to tissue processing than unprocessed tissue analyzed by CLSM
Compared to the Quantum dot (QD) studies performed in our lab, these results show differences between surface coatings and the ability to skin penetration
Summary
Confocal laser scanning microscopy (CLSM) is a classical technique to generate images from cell or tissue samples by means of laser scanning on an optical platform. When the laser is able to penetrate the skin to “resolve” the interior details of the nonopaque specimens, the three dimensional structure of the superficial interior can be constructed. The quality of the images is greatly enhanced due to the unique sectioning method of the laser, and the information from the out-of-focus field is not superimposed on the image in focus. Most of the traditional techniques used to visualize cells or tissues are accompanied by some distortion of the tissue, which may be caused by tissue fixation and sectioning and by duration of exposure to the beams of light. Additional details regarding the mechanisms and methods of CLSM may be readily found on the website of the microscope manufacturer
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