Assessment of Raman hyperspectral spectra upon depletion of proteins involved in iron transport

Pampa Mandal; Alireza Sheikhsofla; Samaneh Ghazanfarpour; Ting Chean Khoo; Jonathan Barra; Isaiah Crousborne; Anna Sharikova; Alexander Khmaladze; Margarida Barroso

doi:10.1117/12.2650402

16 March 2023 Assessment of Raman hyperspectral spectra upon depletion of proteins involved in iron transport

Pampa Mandal, Alireza Sheikhsofla, Samaneh Ghazanfarpour, Ting Chean Khoo, Jonathan Barra, Isaiah Crousborne, Anna Sharikova, Alexander Khmaladze, Margarida Barroso

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Proceedings Volume 12391, Label-free Biomedical Imaging and Sensing (LBIS) 2023; 123910L (2023) https://doi.org/10.1117/12.2650402
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

Raman hyperspectral imaging enables visualization and measurement of the distribution of iron-binding proteins, as well as assessment of the role of proteins involved in the regulation of intracellular iron transport and membrane trafficking in intact cells. To advance the mechanistic understanding of intracellular iron transport, we have performed Raman imaging in MDA-MB-231 wild-type breast cancer cells and compared them to CRISPR/Cas9-based knock-out cells of two intracellular iron homeostasis regulators: the Divalent Metal Transporter 1 (DMT1) and the small GTPase Rab4A. Multivariate singular value decomposition (SVD) analysis showed that the wild type vs. knockout cell populations of both genes could be separated into two distinct groups. Both DMT1 and Rab4A silencing have significant and distinct impacts in a variety of Raman spectra peaks, indicating a strong impact on cell metabolism. Label-free and non-invasive Raman imaging of DMT1 or Rab4A wild-type vs. knockout breast cancer cells should provide important insights into the regulation of intracellular iron homeostasis and cell metabolism in cells and tissues.

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Pampa Mandal, Alireza Sheikhsofla, Samaneh Ghazanfarpour, Ting Chean Khoo, Jonathan Barra, Isaiah Crousborne, Anna Sharikova, Alexander Khmaladze, and Margarida Barroso "Assessment of Raman hyperspectral spectra upon depletion of proteins involved in iron transport", Proc. SPIE 12391, Label-free Biomedical Imaging and Sensing (LBIS) 2023, 123910L (16 March 2023); https://doi.org/10.1117/12.2650402

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