It is a well-established fact that iron metabolism is disrupted in breast cancer cells. Assessment of iron transport and metabolism is necessary to understand molecular mechanism of breast cancer progression. Previously, Raman spectroscopy has been used to measure the Raman spectral profile of iron-bound proteins in breast cancer cells. By harnessing the principle of inelastic scattering of light, Raman spectroscopy offers a powerful, label-free, and nondestructive tool for determination of molecular structures and analysis of chemical bonds. The current study employed a specific experimental approach to capture shifts in the Raman signature of iron-binding proteins, such as transferrin. Focusing on cytoplasmic regions (exclusive of the nucleus) permits improved analysis of iron-binding proteins localized to vesicles present in the cytoplasm. The acquired spectra were subjected to rigorous analysis using singular value decomposition (SVD), a powerful mathematical technique that possesses the ability to reveal underlying trends and enhance biological analysis and interpretation. It involves detecting overlapping frequency patterns in the dataset. By applying SVD to distinguish the Raman spectral profiles of iron-bound transferrin in breast cancer cells, we obtained accurate results that have played a pivotal role in discerning and characterizing the Raman spectral profile of iron-bound transferrin in breast cancer cells.
Cytochrome c, an essential protein integral to the electron transport chain within cellular mitochondria, plays a crucial role in the intricate process of apoptosis, or programmed cell death. An early event in apoptosis involves the release of cytochrome c from the mitochondria's intermembrane space into the cytoplasm. This paper explores the detection of cytochrome c during apoptosis using Raman spectroscopy, with a specific focus on its release from the mitochondria of human microglial cells (HTHμ). Raman spectroscopy, a non-invasive and label-free analytical technique, allows the examination of biomolecular changes based on their chemical properties. Our experimental approach induced apoptosis in HTHμ cells using methamphetamine (METH) and utilized Raman spectroscopy on both control and apoptotic samples. Through the analysis of spectra by singular value decomposition (SVD), which reveals subtle trends and facilitates biological interpretation, distinct spectral features corresponding to cytochrome c were identified. This evidence supports the concept of cytochrome c release from the mitochondria during apoptosis. The label-free nature and high sensitivity of Raman spectroscopy position it as a promising technique for studying apoptosis in biomedical research and contributing to the development of innovative diagnostic approaches for apoptotic-related disorders.
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.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
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.