Toward Raman Subcellular Imaging of Endothelial Dysfunction

Abstract

Multiple diseases are at some point associated with altered endothelial function, and endothelial dysfunction (ED) contributes to their pathophysiology. Biochemical changes of the dysfunctional endothelium are linked to various cellular organelles, including the mitochondria, endoplasmic reticulum, and nucleus, so organelle-specific insight is needed for better understanding of endothelial pathobiology. Raman imaging, which combines chemical specificity with microscopic resolution, has proved to be useful in detecting biochemical changes in ED at the cellular level. However, the detection of spectroscopic markers associated with specific cell organelles, while desirable, cannot easily be achieved by Raman imaging without labeling. This critical review summarizes the current advances in Raman-based analysis of ED, with a focus on a new approach involving molecular Raman reporters that could facilitate the study of biochemical changes in cellular organelles. Finally, imaging techniques based on both conventional spontaneous Raman scattering and the emerging technique of stimulated Raman scattering are discussed.

Towards Raman-Based Screening of Acute Lymphoblastic Leukemia-Type B (B-ALL) Subtypes

“…Therefore, the golden key to successful differentiation probably lies in subtle features within the biochemical frame but probably does not change cancer cell morphology. This suggests that one should look for alternative chemometric methods for data analysis or consider introducing Raman reporters or markers reflecting molecular/biochemical differences/features that may exaggerate the differences between different leukemia subtypes/differentiate the cellular metabolism features of different leukemia subtypes [54]. So far, such reporters are used in targeting specific cellular structures, such as MitoBADY, which accumulates in the mitochondria, or EdU, which bonds with DNA [54].…”

“…This suggests that one should look for alternative chemometric methods for data analysis or consider introducing Raman reporters or markers reflecting molecular/biochemical differences/features that may exaggerate the differences between different leukemia subtypes/differentiate the cellular metabolism features of different leukemia subtypes [54]. So far, such reporters are used in targeting specific cellular structures, such as MitoBADY, which accumulates in the mitochondria, or EdU, which bonds with DNA [54]. Characteristics and proper segregation of common BCP-ALL into subtypes probably require a wide spectrum of immunophenotyping methods, cytogenetics, and molecular diagnostics.…”

Mitokyne: A Ratiometric Raman Probe for Mitochondrial pH

“…The recent advent of commercial vendors of instruments for SRS has accelerated the integration of this spectroscopic method into pharmaceutical and drug discovery research. , The technique can be used to track and monitor the cellular milieu and its response to external stimuli in a spatiotemporal manner, and in combination with chemical probes, it can be used to image the distribution of drugs, − natural products, − sugars, − and lipids. − Raman-active tags have also been metabolically incorporated into nucleic acids and proteins significantly expanding the potential uses of this instrumentation . The majority of probes reported so far for SRS imaging possess constant signal intensity and frequency, which limits their applications to staining. − An exciting yet overlooked area of Raman imaging is the development of reversible sensors which permit the quantification and monitoring of cellular dynamics. This type of probe would be crucial for exploiting the full potential of SRS imaging in chemical biology and pharmaceutical research …”

Raman Thermometry Nanopipettes in Cancer Photothermal Therapy

“…Raman thermometry is considered to have a great potential in replacing the conventional methods because of its versatile application for investigating living systems . Raman spectroscopy is being currently utilized in biomedical and analytical chemistry. , Nanoparticles have been introduced for a trace amount of heavy metal analysis to estimate the temperature …”