Please use this identifier to cite or link to this item: http://hdl.handle.net/10397/43900
Title: Live imaging of cellular internalization of single colloidal particle by combined label-free and fluorescence total internal reflection microscopy
Authors: Byrne, GD
Vllasaliu, D
Falcone, FH
Somekh, MG 
Stolnik, S
Keywords: Clathrin
Endocytosis
Evanescent wave microscopy
Imaging systems
Live cell imaging
Total internal reflection fluorescence
Issue Date: 2015
Publisher: American Chemical Society
Source: Molecular pharmaceutics, 2015, v. 12, no. 11, p. 3862-3870 How to cite?
Journal: Molecular pharmaceutics 
Abstract: In this work we utilize the combination of label-free total internal reflection microscopy and total internal reflectance fluorescence (TIRM/TIRF) microscopy to achieve a simultaneous, live imaging of single, label-free colloidal particle endocytosis by individual cells. The TIRM arm of the microscope enables label free imaging of the colloid and cell membrane features, while the TIRF arm images the dynamics of fluorescent-labeled clathrin (protein involved in endocytosis via clathrin pathway), expressed in transfected 3T3 fibroblasts cells. Using a model polymeric colloid and cells with a fluorescently tagged clathrin endocytosis pathway, we demonstrate that wide field TIRM/TIRF coimaging enables live visualization of the process of colloidal particle interaction with the labeled cell structure, which is valuable for discerning the membrane events and route of colloid internalization by the cell. We further show that 500 nm in diameter model polystyrene colloid associates with clathrin, prior to and during its cellular internalization. This association is not apparent with larger, 1 μm in diameter colloids, indicating an upper particle size limit for clathrin-mediated endocytosis.
URI: http://hdl.handle.net/10397/43900
ISSN: 1543-8384
EISSN: 1543-8392
DOI: 10.1021/acs.molpharmaceut.5b00215
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