Label free imaging of cell‐substrate contacts by holographic total internal reflection microscopy. Issue 9 (2nd November 2016)

• Record Type: Journal Article
• Authors: Mandracchia, Biagio; Gennari, Oriella; Marchesano, Valentina; Paturzo, Melania; Ferraro, Pietro
• Is Part Of: Journal of biophotonics. Volume 10:Issue 9(2017)
• Page Start: 1163
• Page End: 1170
• Publication Date: 2016-11-02
• Subjects: Microscopy [E05.595] -- Microscopy -- Interference [E05.595.513] -- Cell Shape [G04.320] -- Cell Membrane [A11.284.149]; Photonics -- Periodicals; Optical materials -- Periodicals; Optics -- Periodicals; Medical instruments and apparatus -- Periodicals; 621.3605
• DOI: 10.1002/jbio.201600177
• Languages: English
• ISSNs: 1864-063X

Abstract

Abstract : The study of cell adhesion contacts is pivotal to understand cell mechanics and interaction at substrates or chemical and physical stimuli. We designed and built a HoloTIR microscope for label‐free quantitative phase imaging of total internal reflection. Here we show for the first time that HoloTIR is a good choice for label‐free study of focal contacts and of cell/substrate interaction as its sensitivity is enhanced in comparison with standard TIR microscopy. Finally, the simplicity of implementation and relative low cost, due to the requirement of less optical components, make HoloTIR a reasonable alternative, or even an addition, to TIRF microscopy for mapping cell/substratum topography. As a proof of concept, we studied the formation of focal contacts of fibroblasts on three substrates with different levels of affinity for cell adhesion. Abstract : The manuscript presents a novel holographic total internal reflection microscope for label‐free quantitative phase imaging of cell/substrate interaction. This setup is shown to have an enhanced sensitivity in comparison with standard total internal reflection microscopy for distances smaller than 90 nm.