Measuring in vitro cellular uptake of nanoparticles by transmission electron microscopy

Electron Microscopy and Analysis Group Conference 2013

Brown, A.P.^1 ; Brydson, R.M.D.^1 ; Hondow, N.S.^1

Institute for Materials Research, School of Process, Environmental and Materials Engineering, University of Leeds, Leeds LS2 9JT, United Kingdom^1

关键词: Amorphous silica;    Biomedical applications;    Cellular response;    Engineered nanoparticles;    Intracellular compartments;    Research and development;    Sample preparation;    Superparamagnetic iron oxides;   

Others  :  https://iopscience.iop.org/article/10.1088/1742-6596/522/1/012058/pdf
DOI  :  10.1088/1742-6596/522/1/012058

来源: IOP

【 摘 要 】

Biomedical application of engineered nanoparticles (NPs) is a growing area of research and development. Uncertainty remains as to the mode of action of many NP types and TEM is a tool capable of addressing this if used in conjunction with standard cellular response assays. We will demonstrate imaging of thin sections of fixed, plastic embedded cells by analytical TEM to identify: superparamagnetic iron oxide NP translocation into cell compartments such as endosomes; amorphous silica NP penetration through a cell membrane without membrane encapsulation and zinc oxide NP degradation in cell compartments. We will then discuss how the in vitro cellular responses to a dose of NPs exposed to cell lines can be correlated to the internalized dose per cell section noting however that quantification of the latter requires random sampling procedures or correlation to higher throughout techniques to measure a population of whole cells. Similarly, analytical TEM measures of NP degradation within intracellular compartments will require a more appropriate sample preparation such as cryo-fixation.