scholarly journals Development of Single Cell Raman Spectroscopy for Cancer Screening and Therapy Monitoring

2009 ◽  
Author(s):  
James W. Chan
Keyword(s):  
Raman Spectroscopy ◽  
Cancer Screening ◽  
Single Cell ◽  
Therapy Monitoring

scholarly journals Towards automated cancer screening: Label-free classification of fixed cell samples using wavelength modulated Raman spectroscopy

2018 ◽  
Vol 11 (4) ◽  
pp. e201700244 ◽  
Author(s):  
Lana Woolford ◽  
Mingzhou Chen ◽  
Kishan Dholakia ◽  
C. Simon Herrington
Keyword(s):  
Raman Spectroscopy ◽  
Cancer Screening ◽  
Label Free ◽  
Fixed Cell ◽  
Free Classification

scholarly journals Single-cell discrimination based on optical tweezers Raman spectroscopy

2013 ◽  
Vol 58 (21) ◽  
pp. 2594-2600 ◽  
Author(s):  
HongFei Ma ◽  
Yong Zhang ◽  
AnPei Ye
Keyword(s):  
Raman Spectroscopy ◽  
Single Cell ◽  
Optical Tweezers ◽  
Cell Discrimination

Dynamic characterization of drug resistance and heterogeneity of the gastric cancer cell BGC823 using single-cell Raman spectroscopy

The Analyst ◽  
2018 ◽  
Vol 143 (1) ◽  
pp. 164-174 ◽  
Author(s):  
Yong Zhang ◽  
Ludi Jin ◽  
Jingjing Xu ◽  
Yuezhou Yu ◽  
Lin Shen ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Raman Spectroscopy ◽  
Drug Resistance ◽  
Gastric Carcinoma ◽  
Single Cell ◽  
Gastric Cancer Cell ◽  
Carcinoma Cells ◽  
Dynamic Characterization ◽  
Human Gastric Carcinoma Cells

Drug resistance and heterogeneous characteristics of human gastric carcinoma cells (BGC823) under the treatment of paclitaxel (PTX) were investigated using single-cell Raman spectroscopy (RS).

scholarly journals Coarse Raman and optical diffraction tomographic imaging enable label-free phenotyping of isogenic breast cancer cells of varying metastatic potential

2020 ◽  
Author(s):  
Santosh Kumar Paidi ◽  
Vaani Shah ◽  
Piyush Raj ◽  
Kristine Glunde ◽  
Rishikesh Pandey ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Raman Spectroscopy ◽  
Molecular Imaging ◽  
Single Cell ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Metastatic Potential ◽  
Optical Diffraction ◽  
Multivariate Curve Resolution ◽  
Label Free

AbstractIdentification of the metastatic potential represents one of the most important tasks for molecular imaging of cancer. While molecular imaging of metastases has witnessed substantial progress as an area of clinical inquiry, determining precisely what differentiates the metastatic phenotype has proven to be more elusive underscoring the need to marry emerging imaging techniques with tumor biology. In this study, we utilize both the morphological and molecular information provided by 3D optical diffraction tomography and Raman spectroscopy, respectively, to propose a label-free route for optical phenotyping of cancer cells at single-cell resolution. By using an isogenic panel of cell lines derived from MDA-MB-231 breast cancer cells that vary in their metastatic potential, we show that 3D refractive index tomograms can capture subtle morphological differences among the parental, circulating tumor cells, and lung metastatic cells. By leveraging the molecular specificity of Raman spectroscopy, we demonstrate that coarse Raman microscopy is capable of rapidly mapping a sufficient number of cells for training a random forest classifier that can accurately predict the metastatic potential of cells at a single-cell level. We also leverage multivariate curve resolution – alternating least squares decomposition of the spectral dataset to demarcate spectra from cytoplasm and nucleus, and test the feasibility of identifying metastatic phenotypes using the spectra only from the cytoplasmic and nuclear regions. Overall, our study provides a rationale for employing coarse Raman mapping to substantially reduce measurement time thereby enabling the acquisition of reasonably large training datasets that hold the key for label-free single-cell analysis and, consequently, for differentiation of indolent from aggressive phenotypes.

scholarly journals Raman Spectroscopy-Based Measurements of Single-Cell Phenotypic Diversity in Microbial Populations

mSphere ◽  
2020 ◽  
Vol 5 (5) ◽  
Author(s):  
Cristina García-Timermans ◽  
Ruben Props ◽  
Boris Zacchetti ◽  
Myrsini Sakarika ◽  
Frank Delvigne ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Single Cell ◽  
Raman Spectra ◽  
Phenotypic Diversity ◽  
Cell Permeability ◽  
Microbial Populations ◽  
Molecular Profile ◽  
Microbial Cells ◽  
Content Type ◽  
Stressed Cells

ABSTRACT Microbial cells experience physiological changes due to environmental change, such as pH and temperature, the release of bactericidal agents, or nutrient limitation. This has been shown to affect community assembly and physiological processes (e.g., stress tolerance, virulence, or cellular metabolic activity). Metabolic stress is typically quantified by measuring community phenotypic properties such as biomass growth, reactive oxygen species, or cell permeability. However, bulk community measurements do not take into account single-cell phenotypic diversity, which is important for a better understanding and the subsequent management of microbial populations. Raman spectroscopy is a nondestructive alternative that provides detailed information on the biochemical makeup of each individual cell. Here, we introduce a method for describing single-cell phenotypic diversity using the Hill diversity framework of Raman spectra. Using the biomolecular profile of individual cells, we obtained a metric to compare cellular states and used it to study stress-induced changes. First, in two Escherichia coli populations either treated with ethanol or nontreated and then in two Saccharomyces cerevisiae subpopulations with either high or low expression of a stress reporter. In both cases, we were able to quantify single-cell phenotypic diversity and to discriminate metabolically stressed cells using a clustering algorithm. We also described how the lipid, protein, and nucleic acid compositions changed after the exposure to the stressor using information from the Raman spectra. Our results show that Raman spectroscopy delivers the necessary resolution to quantify phenotypic diversity within individual cells and that this information can be used to study stress-driven metabolic diversity in microbial populations. IMPORTANCE Microbial cells that live in the same community can exist in different physiological and morphological states that change as a function of spatiotemporal variations in environmental conditions. This phenomenon is commonly known as phenotypic heterogeneity and/or diversity. Measuring this plethora of cellular expressions is needed to better understand and manage microbial processes. However, most tools to study phenotypic diversity only average the behavior of the sampled community. In this work, we present a way to quantify the phenotypic diversity of microbial samples by inferring the (bio)molecular profile of its constituent cells using Raman spectroscopy. We demonstrate how this tool can be used to quantify the phenotypic diversity that arises after the exposure of microbes to stress. Raman spectroscopy holds potential for the detection of stressed cells in bioproduction.

scholarly journals Proteorhodopsin Overproduction Enhances the Long-Term Viability of Escherichia coli

2019 ◽  
Vol 86 (1) ◽  
Author(s):  
Yizhi Song ◽  
Michaël L. Cartron ◽  
Philip J. Jackson ◽  
Paul A. Davison ◽  
Mark J. Dickman ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Raman Spectroscopy ◽  
Cell Membrane ◽  
Single Cell ◽  
Marine Bacteria ◽  
Membrane Area ◽  
Content Type ◽  
E Coli ◽  
Wide Range

ABSTRACT Genes encoding the photoreactive protein proteorhodopsin (PR) have been found in a wide range of marine bacterial species, reflecting the significant contribution that PR makes to energy flux and carbon cycling in ocean ecosystems. PR can also confer advantages to enhance the ability of marine bacteria to survive periods of starvation. Here, we investigate the effect of heterologously produced PR on the viability of Escherichia coli. Quantitative mass spectrometry shows that E. coli, exogenously supplied with the retinal cofactor, assembles as many as 187,000 holo-PR molecules per cell, accounting for approximately 47% of the membrane area; even cells with no retinal synthesize ∼148,000 apo-PR molecules per cell. We show that populations of E. coli cells containing PR exhibit significantly extended viability over many weeks, and we use single-cell Raman spectroscopy (SCRS) to detect holo-PR in 9-month-old cells. SCRS shows that such cells, even incubated in the dark and therefore with inactive PR, maintain cellular levels of DNA and RNA and avoid deterioration of the cytoplasmic membrane, a likely basis for extended viability. The substantial proportion of the E. coli membrane required to accommodate high levels of PR likely fosters extensive intermolecular contacts, suggested to physically stabilize the cell membrane and impart a long-term benefit manifested as extended viability in the dark. We propose that marine bacteria could benefit similarly from a high PR content, with a stabilized cell membrane extending survival when those bacteria experience periods of severe nutrient or light limitation in the oceans. IMPORTANCE Proteorhodopsin (PR) is part of a diverse, abundant, and widespread superfamily of photoreactive proteins, the microbial rhodopsins. PR, a light-driven proton pump, enhances the ability of the marine bacterium Vibrio strain AND4 to survive and recover from periods of starvation, and heterologously produced PR extends the viability of nutrient-limited Shewanella oneidensis. We show that heterologously produced PR enhances the viability of E. coli cultures over long periods of several weeks and use single-cell Raman spectroscopy (SCRS) to detect PR in 9-month-old cells. We identify a densely packed and consequently stabilized cell membrane as the likely basis for extended viability. Similar considerations are suggested to apply to marine bacteria, for which high PR levels represent a significant investment in scarce metabolic resources. PR-stabilized cell membranes in marine bacteria are proposed to keep a population viable during extended periods of light or nutrient limitation, until conditions improve.

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