Tracking nanoparticles in three-dimensional tissue-engineered models using confocal laser scanning microscopy.
Here we describe a method for imaging the position of nanoparticles within a 3D tissue-engineered model using confocal laser scanning microscopy (CLSM). The ability to track diffusion of nanoparticles in vitro is an important part of trans-dermal and trans-mucosal drug delivery development as well as for intra-epithelial drug delivery. Using 3D tissue-engineered models enables us to image diffusion in vitro in a physiologically relevant way; not possible in two-dimensional monolayer cultures (MacNeil, Nature 445:874-880, 2007; Hearnden et al., Pharmaceutical Res. 26(7):1718-1728, 2009). CLSM enables imaging of viable in vitro models in three dimensions with good spatial and axial resolution (Georgakoudi et al., Tissue Eng 14:1-20, 2008; Schenke-Layland et al., Adv. Drug Del. Rev. 58:878-896, 2006). Here we show that fluorescently labelled nanoparticles can be visualised, quantified, and their position within the cell can be determined using CLSM.
Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield, UK.
Vanessa
V
Hearnden
Sheila
S
MacNeil
Giuseppe
G
Battaglia
D000818:Animals; D004058:Diffusion; D005456:Fluorescent Dyes / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D018613:Microscopy, Confocal / Q000379:methods*; D008954:Models, Biological; D053758:Nanoparticles / Q000737:chemistry*; D011108:Polymers / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry
D005456:Fluorescent Dyes; D011108:Polymers; D012235:Rhodamines