Investigation on cytoskeleton dynamics for non-adherent cells under point-like stimuli

Lisa Miccio; Pasquale Memmolo; Francesco Merola; Martina Mugnano; Sabato Fusco; Antonio Paciello; Pietro Ferraro; Paolo A. Netti

doi:10.1117/12.2185929

22 June 2015 Investigation on cytoskeleton dynamics for non-adherent cells under point-like stimuli

Lisa Miccio, Pasquale Memmolo, Francesco Merola, Martina Mugnano, Sabato Fusco, Antonio Paciello, Pietro Ferraro, Paolo A. Netti

Author Affiliations +

Proceedings Volume 9529, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials II; 95290P (2015) https://doi.org/10.1117/12.2185929
Event: SPIE Optical Metrology, 2015, Munich, Germany

Abstract

In the present paper, Holographic Optical Tweezers (HOT) is employed to trap and manage functionalized micrometric latex beads with the aim at probing cellular forces in no-adherent state. For the first time at best of our knowledge, a suspended cell, subjected to mechanical stress, structures its cytoskeleton when anchored to point-like bonds. We exploit the HOT arrangement to induce mechanical deformation in suspended NIH 3T3 fibroblast. Our investigation is devoted to understand the inner cell mechanism when it is mechanically stressed by point-like stimulus without the substrate influence. In our experiment, cell adhesion is prevented and the stimulus is applied through latex beads trapped by HOT and positioned externally to the cell membrane. Our aims are devoted to analyze cell response during the transition from an homogeneous and isotropic structure (as it’s in suspension) to a mechanically stressed state. To analyze the cell material interaction we combine the HOT arrangement with two imaging systems: a Digital Holography (DH) setup in microscope configuration that is an investigation method useful for quantitative, label-free and full-field analysis of low contrast object and a fluorescence modulus. HOT are exploited to induce cellular response to specific stimuli while DH allows to measure such responses in no-invasive way. Finally, fluorescence imaging is added to discriminate the inner cell structures.

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Lisa Miccio, Pasquale Memmolo, Francesco Merola, Martina Mugnano, Sabato Fusco, Antonio Paciello, Pietro Ferraro, and Paolo A. Netti "Investigation on cytoskeleton dynamics for non-adherent cells under point-like stimuli", Proc. SPIE 9529, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials II, 95290P (22 June 2015); https://doi.org/10.1117/12.2185929

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