High-fidelity probing of the structure and heterogeneity of extracellular vesicles by resonance-enhanced atomic force microscopy infrared spectroscopy

Kim, Sally Yunsun; Khanal, Dipesh; Kalionis, Bill; Chrzanowski, Wojciech

doi:10.1038/s41596-018-0109-3

Cited by 71 publications

(65 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, OMV heights in AFM images were a fraction of the OMV width, only 5-15 nm (data not shown), implying that the OMVs flattened during sample desiccation and these diameters may not represent true in vivo OMV size. These size ranges and this potential vesicle flattening has been observed by other researchers imaging vesicles with AFM 67 .…”

Section: Resultssupporting

confidence: 77%

“…DLS and AFM measurements showed that OMV sizes were highly polydisperse in size regardless of the cellular stressor. Differing OMV diameter according to the analysis method has been reported in the literature 67 , likely due to differences in OMV aggregation and/or hydration. Aggregation of OMVs observed in AFM and SEM images may be due to outer membrane surface features similar to those involved in adhesion for intact cells 69 .…”

Section: Discussionmentioning

confidence: 83%

See 1 more Smart Citation

Abiotic stressors impact outer membrane vesicle composition in a beneficial rhizobacterium: Raman spectroscopy characterization

Potter

Hanson

Anderson

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Outer membrane vesicles (OMVs) produced by Gram-negative bacteria have roles in cell-to-cell signaling, biofilm formation, and stress responses. Here, the effects of abiotic stressors on OMV contents and composition from biofilm cells of the plant health-promoting bacterium Pseudomonas chlororaphis O6 (PcO6) are examined. Two stressors relevant to this root-colonizing bacterium were examined: CuO nanoparticles (NPs)-a potential fertilizer and fungicide- and H2O2-released from roots during plant stress responses. Atomic force microscopy revealed 40–300 nm diameter OMVs from control and stressed biofilm cells. Raman spectroscopy with linear discriminant analysis (LDA) was used to identify changes in chemical profiles of PcO6 cells and resultant OMVs according to the cellular stressor with 84.7% and 83.3% accuracies, respectively. All OMVs had higher relative concentrations of proteins, lipids, and nucleic acids than PcO6 cells. The nucleic acid concentration in OMVs exhibited a cellular stressor-dependent increase: CuO NP-induced OMVs > H2O2-induced OMVs > control OMVs. Biochemical assays confirmed the presence of lipopolysaccharides, nucleic acids, and protein in OMVs; however, these assays did not discriminate OMV composition according to the cellular stressor. These results demonstrate the sensitivity of Raman spectroscopy using LDA to characterize and distinguish cellular stress effects on OMVs composition and contents.

show abstract

Section: Resultssupporting

confidence: 77%

Section: Discussionmentioning

confidence: 83%

Abiotic stressors impact outer membrane vesicle composition in a beneficial rhizobacterium: Raman spectroscopy characterization

Potter

Hanson

Anderson

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…A decrease in immunosuppressive exosomes associated with preterm birth and shifts of molecular cargo towards pro-coagulant and pro-inflammatory factors in trophoblast-derived EVs in preeclampsia are suggestive of their contribution to a modulated maternal immune adaptation in complicated pregnancies, although causation has not been demonstrated [ 114 ]. Shortcomings in resolution and sensitivity of techniques to quantitatively and qualitatively assess nano-scale EVs have so far been an obstacle to the understanding of the biogenesis and activity of their various forms; however, a number of new technologies, such as combined differential ultracentrifugation, transmission electron microscopy, and nanoparticle tracking analysis have emerged that allow for their isolation, examination of morphology, and analysis of size distribution and concentration [ 78 , 113 , 193 ].…”

Section: Feto-placental Pacemakers Programming the Immune Clock Of Prmentioning

confidence: 99%

Multiomic immune clockworks of pregnancy

et al. 2020

View full text Add to dashboard Cite

Preterm birth is the leading cause of mortality in children under the age of five worldwide. Despite major efforts, we still lack the ability to accurately predict and effectively prevent preterm birth. While multiple factors contribute to preterm labor, dysregulations of immunological adaptations required for the maintenance of a healthy pregnancy is at its pathophysiological core. Consequently, a precise understanding of these chronologically paced immune adaptations and of the biological pacemakers that synchronize the pregnancy "immune clock" is a critical first step towards identifying deviations that are hallmarks of peterm birth. Here, we will review key elements of the fetal, placental, and maternal pacemakers that program the immune clock of pregnancy. We will then emphasize multiomic studies that enable a more integrated view of pregnancy-related immune adaptations. Such multiomic assessments can strengthen the biological plausibility of immunological findings and increase the power of biological signatures predictive of preterm birth

show abstract

“…This drawback is often circumvented by coupling the analytes with plasmonic nanoparticles (SERS) [15,16], by labelling them with organic dyes (Raman reporters), or immune-labelling with nanoprobes [17]. These strategies allowed to distinguish EVs from various sources, disease contexts [18,19] and singlevesicle analysis [20][21][22]. SERS-active nanostructures can enhance the Raman signals of target molecules by several orders of magnitude, but can have problems in stability, poor reproducibility, and sample damaging according to their material composition and architecture [23,24].…”

Section: Introductionmentioning

confidence: 99%

Fourier‐transform Infrared (FT‐IR) spectroscopy fingerprints subpopulations of extracellular vesicles of different sizes and cellular origin

Paolini

Federici

Consoli

et al. 2020

J of Extracellular Vesicle

View full text Add to dashboard Cite

2020) Fourier-transform Infrared (FT-IR) spectroscopy fingerprints subpopulations of extracellular vesicles of different sizes and cellular ABSTRACT Identification of extracellular vesicle (EV) subpopulations remains an open challenge. To date, the common strategy is based on searching and probing set of molecular components and physical properties intended to be univocally characteristics of the target subpopulation. Pitfalls include the risk to opt for an unsuitable marker setwhich may either not represent the subpopulation or also cover other unintended subpopulationsand the need to use different characterization techniques and equipment. This approach focused on specific markers may result inadequate to routinely deal with EV subpopulations that have an intrinsic high level of heterogeneity. In this paper, we show that Fourier-transform Infrared (FT-IR) spectroscopy can provide a collective fingerprint of EV subpopulations in one single experiment. FT-IR measurements were performed on large (LEVs,~600 nm), medium (MEVs,~200 nm) and small (SEVs~60 nm) EVs enriched from two different cell lines medium: murine prostate cancer (TRAMP-C2) and skin melanoma (B16). Spectral regions between 3100-2800 cm −1 and 1880-900 cm −1 , corresponding to functional groups mainly ascribed to lipid and protein contributions, were acquired and processed by Principal Component Analysis (PCA). LEVs, MEVs and SEVs were separately grouped for both the considered cell lines. Moreover, subpopulations of the same size but from different sources were assigned (with different degrees of accuracy) to two different groups. These findings demonstrate that FT-IR has the potential to quickly fingerprint EV subpopulations as a whole, suggesting an appealing complement/alternative for their characterization and grading, extendable to healthy and pathological EVs and fully artificial nanovesicles. ARTICLE HISTORY

show abstract

High-fidelity probing of the structure and heterogeneity of extracellular vesicles by resonance-enhanced atomic force microscopy infrared spectroscopy

Cited by 71 publications

References 45 publications

Abiotic stressors impact outer membrane vesicle composition in a beneficial rhizobacterium: Raman spectroscopy characterization

Abiotic stressors impact outer membrane vesicle composition in a beneficial rhizobacterium: Raman spectroscopy characterization

Multiomic immune clockworks of pregnancy

Fourier‐transform Infrared (FT‐IR) spectroscopy fingerprints subpopulations of extracellular vesicles of different sizes and cellular origin

Contact Info

Product

Resources

About