Detection of cells with lipid droplets exploiting their biolens’ signature in holographic imaging flow cytometry

Daniele Pirone; Daniele G. Sirico; Martina Mugnano; Danila Del Giudice; Ivana Kurelac; Beatrice Cavina; Pasquale Memmolo; Lisa Miccio; Pietro Ferraro

doi:10.1117/12.2674987

11 August 2023 Detection of cells with lipid droplets exploiting their biolens’ signature in holographic imaging flow cytometry

Daniele Pirone, Daniele G. Sirico, Martina Mugnano, Danila Del Giudice, Ivana Kurelac, Beatrice Cavina, Pasquale Memmolo, Lisa Miccio, Pietro Ferraro

Author Affiliations +

Proceedings Volume 12622, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI; 1262218 (2023) https://doi.org/10.1117/12.2674987
Event: SPIE Optical Metrology, 2023, Munich, Germany

Conference Poster

Abstract

In recent years, the dynamic role of Lipid Droplets (LDs) in many cellular activities has been increasingly brought to light. In fact, it has been discovered that LDs are involved in many pathologies (e.g., diabetes, atherosclerosis, pathogen infections, neurodegenerative diseases and cancer). Moreover, it has been demonstrated that their number and size increase during an inflammation or infectious inside the immune cells, also with the COVID-19. Therefore, detecting LDs within single cells could aid the diagnosis of several pathologies. Currently, the gold-standard technique in this field is Fluorescence Imaging Flow Cytometry (FIFC), in which the single-cell analysis of fluorescence microscopy is implemented in high-throughput modality thanks to the flow-cytometry module. However, to overcome the drawbacks related to the fluorescence staining, Holographic Imaging Flow Cytometry (HIFC) has gaining momentum as label-free alternative to the FIFC tool. Thanks to the interferometric principles at the basis of digital holography, it has been already demonstrated that a suspended cell acts as a biological lens with specific focusing features. Here we show that the presence of intracellular LDs inside the cell is able to change its focalization features, measured through a HIFC system. Therefore, based on this property, we demonstrate that a detection of single cells containing intracellular LDs is possible by means of a direct analysis of the digital holograms recorded in flow cytometry modality. The attained results open the route to the development of a fast, non-destructive, and high-throughput tool for the diagnosis of LDs-related pathologies by exploiting the biolens’ signature in HIFC.

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Daniele Pirone, Daniele G. Sirico, Martina Mugnano, Danila Del Giudice, Ivana Kurelac, Beatrice Cavina, Pasquale Memmolo, Lisa Miccio, and Pietro Ferraro "Detection of cells with lipid droplets exploiting their biolens’ signature in holographic imaging flow cytometry", Proc. SPIE 12622, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI, 1262218 (11 August 2023); https://doi.org/10.1117/12.2674987

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