Fig. 2: PmBF Specifically Recognizes and Quantitatively Detects OMVs. | Nature Communications

Fig. 2: PmBF Specifically Recognizes and Quantitatively Detects OMVs.

From: Specific labeling of outer membrane vesicles with antibiotic-conjugated probe reveals early bacterial infections in blood

Fig. 2

a Super-resolution images of Gram-negative bacteria, Gram-positive bacteria and mammal cell after incubation with PmBF for 1 h. b Fluorescence intensity of each group were detected by cytation 5, RFU: Relative Fluorescence Units (Bars represent the mean ± SD, n = 3 biological replicates). c Nanoparticles tracking analysis (NTA) of E. coli OMVs. d Western blot images of E. coli and OMVs, the protein loading amount was 50 μg for the bacterial samples and 20 μg for the EVs sample. e Transmission electron microscopy (TEM) images of E. coli OMVs. f Super-resolution images of E. coli OMVs, HeLa EVs and S. aureus EVs labeled with CellMask™ Deep Red plasma membrane stain (red) and PmBF (green). g Schematic representation of nano-flow cytometry (Created with BioRender.com). h, i Natural extracellular vesicles (control group) and debris sample detection signal and concentrations detected based on nano-flow cytometry (Bars represent the mean ± SD, n = 3 biological replicates). j Quantitative analysis of E. coli OMVs with different concentration gradients by nano-flow cytometry. k Quantitative analysis of different vesicles after PmBF labelling by nano-flow cytometry (Bars represent the mean ± SD, n = 3 biological replicates). l Quantitative analysis of positive VLDL after labeling by PmBF or antibody probes, control group was treated with PBS (Bars represent the mean ± SD, n = 3 biological replicates). m PmBF probe was used to detect E. coli OMVs within a mixture of various vesicle types (S. aureus and HeLa EVs). Group HeLa: 100% HeLa EVs; Group S. aureus: 100% S. aureus EVs; Group EHS: 40% E.coli OMVs + 30% HeLa EVs + 30% S. aureus EVs; Group EH: 50% E.coli OMVs + 50% HeLa EVs; Group ES: 50% E.coli OMVs + 50% S. aureus EVs; Group E. coli: 100% E.coli OMVs (Bars represent the mean ± SD, n = 3 biological replicates). b, k, l were determined by one-way ANOVA with multiplicity adjusted P value; i was determined by a two-tailed unpaired t-test (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns: P ≥ 0.05). Source data are provided as a Source Data file.

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