Ivan Kempson - Quantitative analysis of nanoparticle internalization in mammalian cells by high resolution X-ray microscopy

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      Publication Details (including relevant citation   information):

      HH Chen, CC   Chien, C Petibois, CL Wang, YS Chu, SF Lai, TE Hua, YY Chen, X   Cai, IM Kempson, Y Hwu, G Margaritondo, Quantitative analysis of   nanoparticle internalization in mammalian cells by high   resolution X-ray microscopy, Journal of   Nanobiotechnology, 9, art no 14. DOI:   10.1186/1477-3155-9-14, 2011.


      Background: Quantitative analysis of nanoparticle uptake at the   cellular level is critical to nanomedicine procedures. In   particular, it is required for a realistic evaluation of their   effects. Unfortunately, quantitative measurements of nanoparticle   uptake still pose a formidable technical challenge. We present   here a method to tackle this problem and analyze the number of   metal nanoparticles present in different types of cells. The   method relies on high-lateral-resolution (better than 30 nm)   transmission x-ray microimages with both absorption contrast and   phase contrast – including two-dimensional (2D) projection images   and three-dimensional (3D) tomographic reconstructions that   directly show the nanoparticles.

      Results: Practical tests were successfully conducted on bare and   polyethylene glycol (PEG) coated gold nanoparticles obtained by   x-ray irradiation. Using two different cell lines, EMT and HeLa,   we obtained the number of nanoparticle clusters uptaken by each   cell and the cluster size. Furthermore, the analysis revealed   interesting differences between 2D and 3D cultured cells as well   as between 2D and 3D data for the same 3D specimen.

      Conclusions: We demonstrated the feasibility and effectiveness of   our method, proving that it is accurate enough to measure the   nanoparticle uptake differences between cells as well as the   sizes of the formed nanoparticle clusters. The differences   between 2D and 3D cultures and 2D and 3D images stress the   importance of the 3D analysis which is made possible by our   approach.

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