Differences between wild type and LSD populations of HeLa and MEF cells through investigation of lysosomal aggregation and morphological variations by digital holography

Giusy Giugliano; Jaromir Behal; Daniele Pirone; Michela Schiavo; Antonella Capuozzo; Sandro Montefusco; Vittorio Bianco; Lisa Miccio; Pasquale Memmolo; Diego Medina; Pietro Ferraro

doi:10.1117/12.2675432

11 August 2023 Differences between wild type and LSD populations of HeLa and MEF cells through investigation of lysosomal aggregation and morphological variations by digital holography

Giusy Giugliano, Jaromir Behal, Daniele Pirone, Michela Schiavo, Antonella Capuozzo, Sandro Montefusco, Vittorio Bianco, Lisa Miccio, Pasquale Memmolo, Diego Medina, Pietro Ferraro

Author Affiliations +

Proceedings Volume 12622, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI; 126220M (2023) https://doi.org/10.1117/12.2675432
Event: SPIE Optical Metrology, 2023, Munich, Germany

Abstract

The most used imaging method for biological samples is based on the use of markers or fluorescent colorants to label the cells micro-structures of interest. However, labeling the cell may cause alteration of its internal constituents, its natural behavior and its life cycle in the case of living cells. Digital Holography (DH) in microscopy is a powerful imaging technique which permits to obtain a posteriori multiple refocusing and quantitative phase contrast images. The main advantage of the DH is the ability to provide the cell morphological features in label-free mode. DH has been proved successfully in different biomedical applications, such as characterization and identification of cancer cells, diagnosis of blood diseases and marker-free detection of lipid droplets. We implemented a Mach-Zehnder interferometer in off-axis configuration which allows recording the resultant digital holograms. Therefore, we performed the 2D numerical reconstruction to achieve the quantitative phase maps through several computational steps, namely Fourier spectrum filtering, numerical refocusing, aberrations suppression and phase unwrapping. Here, we show a detailed study of two different classes of biological samples: HeLa cells and mouse embryonic fibroblasts. Specially, through the proposed method, we investigate the morphological variations induced by lysosomal aggregations to distinguish the difference between lysosomal storage diseases and wild type populations of both cell lines. This work demonstrates the validity and effectiveness of the presented method, revealing its potential to discriminate between healthy and unhealthy cells at subcellular level.

Conference Presentation

Citation Download Citation

Giusy Giugliano, Jaromir Behal, Daniele Pirone, Michela Schiavo, Antonella Capuozzo, Sandro Montefusco, Vittorio Bianco, Lisa Miccio, Pasquale Memmolo, Diego Medina, and Pietro Ferraro "Differences between wild type and LSD populations of HeLa and MEF cells through investigation of lysosomal aggregation and morphological variations by digital holography", Proc. SPIE 12622, Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI, 126220M (11 August 2023); https://doi.org/10.1117/12.2675432

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available