Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Optical Diffraction Tomography and Raman Confocal Microscopy for the Investigation of Vacuoles Associated with Cancer Senescent Engulfing Cells

Silvia Ghislanzoni
* ,
Jeon Woong Kang
* ORCID logo ,
Arianna Bresci
,
Andrea Masella
ORCID logo ,
Koseki J. Kobayashi-Kirschvink
,
Dario Polli
ORCID logo ,
Italia Bongarzone
† ,
Peter T. C. So
†
†
These authors have contributed equally to this work.
Version 1 : Received: 9 October 2023 / Approved: 10 October 2023 / Online: 10 October 2023 (05:32:15 CEST)

A peer-reviewed article of this Preprint also exists.

Ghislanzoni, S.; Kang, J.W.; Bresci, A.; Masella, A.; Kobayashi-Kirschvink, K.J.; Polli, D.; Bongarzone, I.; So, P.T.C. Optical Diffraction Tomography and Raman Confocal Microscopy for the Investigation of Vacuoles Associated with Cancer Senescent Engulfing Cells. Biosensors 2023, 13, 973. Ghislanzoni, S.; Kang, J.W.; Bresci, A.; Masella, A.; Kobayashi-Kirschvink, K.J.; Polli, D.; Bongarzone, I.; So, P.T.C. Optical Diffraction Tomography and Raman Confocal Microscopy for the Investigation of Vacuoles Associated with Cancer Senescent Engulfing Cells. Biosensors 2023, 13, 973.

Abstract

Wild-type p53 cancer therapy-induced senescent cells frequently engulf and degrade neighboring ones inside a massive vacuole in their cytoplasm. After clearance of the internalized cell, the vacuole persists, seemingly empty, for several hours. Despite large vacuoles being associated with cell death, this process is known to confer a survival advantage to cancer engulfing cells, leading to therapy resistance and tumor relapse. Previous attempts to resolve the vacuolar structure and visualize their content using dyes were unsatisfying for lack of known targets and ineffective dye penetration and/or retain. Here, we overcame this problem by applying optical diffraction tomography and Raman spectroscopy to MCF7 doxorubicin-induced engulfing cells. We demonstrated a real ability of cell tomography and Raman to phenotype complex microstructures, such as cell-in-cells and vacuoles, and detect chemical species in extremely low concentrations within live cells in a completely label-free fashion. We show that vacuoles had a density indistinguishable to the medium, but were not empty, instead contained diluted cell-derived macromolecules, and we could discern vacuoles from medium and cells using their Raman fingerprint. Our approach is useful for the non-invasive investigation of senescent engulfing (and other peculiar) cells in unperturbed conditions, crucial for a better understanding of complex biological processes.

Keywords

microscopy; optical diffraction tomography; Raman spectroscopy; therapy-induced senescence; cell engulfing; vacuole; cancer; cell-in-cell

Subject

Biology and Life Sciences, Life Sciences

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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