Journal: RSC Advances

Article Title: Development of a platform for quantum-dot-labeled polyethylene nanoplastics for dynamic analytical applications

doi: 10.1039/d6ra01781a

Figure Lengend Snippet: Scanning emission microscopy (SEM) images and particle size distribution and zeta potential of the prepared particles. SEM images of unlabeled polyethylene nanoplastics (nPE) (A), and nPE utilizing Qdots (B and C). Scale bars: 1 µm. SEM images were analysed using ImageJ software. Feret diameter (D) and roundness (E) of plastic particles are shown. (F) Zeta potential of unlabeled polyethylene nanoplastics (nPE), green fluorescent labeled nPE (1 mg mL −1 ) in milliQ containing 0.05% tween are shown.

Article Snippet: Fluorescent-labeled nPE was imaged using CellVoyager CV8000 (Yokogawa, Tokyo, Japan) with 405 nm excitation and 525/50 nm emission filters for green Qdots, and 405 nm excitation and 600/37 nm emission filters for red Qdots.

Techniques: Microscopy, Zeta Potential Analyzer, Software, Labeling

Journal: RSC Advances

Article Title: Development of a platform for quantum-dot-labeled polyethylene nanoplastics for dynamic analytical applications

doi: 10.1039/d6ra01781a

Figure Lengend Snippet: THP-1 cells (2.0 × 104 cells per well) were seeded in 96 well plates and differentiated with PMA for 24 h. Cells were then exposed to unlabeled nPE, green fluorescent labeled nPE and red fluorescent labeled nPE (0, 0.1, 1.0, or 10 mg mL −1 in 0.1% CMC medium) under continuous shaking for 24 h, followed by MTT assay. Data represent from two independent experiments.

Article Snippet: Fluorescent-labeled nPE was imaged using CellVoyager CV8000 (Yokogawa, Tokyo, Japan) with 405 nm excitation and 525/50 nm emission filters for green Qdots, and 405 nm excitation and 600/37 nm emission filters for red Qdots.

Techniques: Labeling, MTT Assay

Journal: RSC Advances

Article Title: Development of a platform for quantum-dot-labeled polyethylene nanoplastics for dynamic analytical applications

doi: 10.1039/d6ra01781a

Figure Lengend Snippet: Physicochemical properties of degraded labelled nPE. (A) SEM images of red fluorescent labeled degraded nPE (Red d-nPE). Scale bars: 3 µm. Feret diameter (B) and roundness (C) of red d-nPE particles calculated through SEM image analysis. (D) Relative ATR-IR spectra of Red d-nPE. (E) Zeta potential of Red d-nPE (1 mg mL −1 ) in milliQ containing 0.05% tween. (F) Measurement of fluorescence intensity of 0.25, 0.5, 1 mg mL −1 red d-nPE in EtOH using a plate reader. These experiments were repeated twice with similar results. Note that significance was assessed in using one way ANOVA followed by Dunnett's multiple comparison test as follows: *, P < 0.05; ***, P < 0.001. (G) THP-1 cells (2.0 × 10 4 cells per well) were seeded in 96 well plates and differentiated with PMA for 24 h. Cells were then exposed to red d-nPE (0, 0.1, 1.0, or 10 mg mL −1 in 0.1% CMC medium) under continuous shaking for 24 h, followed by MTT assay. Data represent from two independent experiments.

Article Snippet: Fluorescent-labeled nPE was imaged using CellVoyager CV8000 (Yokogawa, Tokyo, Japan) with 405 nm excitation and 525/50 nm emission filters for green Qdots, and 405 nm excitation and 600/37 nm emission filters for red Qdots.

Techniques: Labeling, Zeta Potential Analyzer, Fluorescence, Comparison, MTT Assay

Journal: RSC Advances

Article Title: Development of a platform for quantum-dot-labeled polyethylene nanoplastics for dynamic analytical applications

doi: 10.1039/d6ra01781a

Figure Lengend Snippet: Cellular uptake of labelled nPE. (A) THP-1 cells (2.0 × 10 4 cells per well) were seeded in 96 well plates and differentiated with PMA for 24 h. Cells were then exposed to red fluorescent labeled nPE and red fluorescent labeled d-nPE (1.0 mg mL −1 in 0.1% CMC medium) under continuous shaking for 24 h at 37 °C. (B) The proportion of cells with adherent/incorporated nPE and d-nPE was calculated from the obtained fluorescence images. These experiments were repeated twice with similar results. Note that significance was assessed in using one way ANOVA followed by Tukey's test as follows: ****, P < 0.0001.

Article Snippet: Fluorescent-labeled nPE was imaged using CellVoyager CV8000 (Yokogawa, Tokyo, Japan) with 405 nm excitation and 525/50 nm emission filters for green Qdots, and 405 nm excitation and 600/37 nm emission filters for red Qdots.

Techniques: Labeling, Fluorescence

Journal: bioRxiv

Article Title: Sequence-Independent In Situ Imaging of GlycoRNA in Living Cells and Tissues based on COMPASS

doi: 10.64898/2026.04.01.715772

Figure Lengend Snippet: a, CLSM images of glycoRNAs on the surface of MCF-7 cells with COMPASS and various control groups lacking components of the COMPASS. w/o, without. Scale bar, 100 μm. b, Flow cytometric analysis of COMPASS-processed MCF-7 cells incubated with Ac 4 ManNAz, 5-EU, or both, followed by sequential treatment with DBCO-AF647 and N 3 -AF488. c, RT-qPCR analysis of RNA samples extracted from DTWD2 gene silencing by siRNA and the negative control (NC). d, CLSM images of MCF-7 cells after silencing of the DTWD2 gene by siRNA or NC. Scale bar, 20 μm. e, Quantification of average fluorescence intensity in d . Data in e is representative of three independent experiments; n = 5 frames. f, Colocalization images (overlay channel) of glycoRNAs (COMPASS channel) with plasma membrane (DiD channel) or lipid rafts (CT-B channel). Scale bar, 10 μm. The line diagram depicts the intensity profiles of the fluorescent signals along the yellow arrows in the overlay images. The Pearson scatterplot on the right illustrates the degree of colocalization in the overlay image. Data are shown as mean ± s.d. Statistical significance was analyzed by unpaired t-test; NS, not significant;(*) P < 0.05, (**) P < 0.01, (***) P < 0.001, and (****) P < 0.0001.

Article Snippet: Subsequently, DMB labeling was performed with the Sialic Acid Fluorescence Labeling Kit (Takara) and analyzed by HPLC with fluorescence detection.

Techniques: Control, Incubation, Quantitative RT-PCR, Negative Control, Fluorescence, Clinical Proteomics, Membrane

Journal: bioRxiv

Article Title: Sequence-Independent In Situ Imaging of GlycoRNA in Living Cells and Tissues based on COMPASS

doi: 10.64898/2026.04.01.715772

Figure Lengend Snippet: a, Specificity for COMPASS to image glycoRNAs on the MCF-7 cell surface. Imaging with COMPASS after treating MCF-7 cells with two RNases (RNase A and RNase T1), two glycosidases (PNGase F and NA), two glycosylation inhibitors (tunicamycin and BG), or two proteases (trypsin and proteinase K), respectively. Scale bar, 20 μm. b, Quantification of average fluorescence intensity in a . Data in b is representative of three independent experiments; n = 5 frames. Data are shown as mean ± s.d. Statistical significance was analyzed by unpaired two-tailed Student’s t-test; NS, not significant;(*) P < 0.05, (**) P < 0.01, (***) P < 0.001, and (****) P < 0.0001. c, Generality for COMPASS to image glycoRNAs on the cell surface of Hela cells, HepG2 cells, LM3 cells, and IMR-32 cells. Scale bar, 20 μm.

Article Snippet: Subsequently, DMB labeling was performed with the Sialic Acid Fluorescence Labeling Kit (Takara) and analyzed by HPLC with fluorescence detection.

Techniques: Imaging, Glycoproteomics, Fluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Sequence-Independent In Situ Imaging of GlycoRNA in Living Cells and Tissues based on COMPASS

doi: 10.64898/2026.04.01.715772

Figure Lengend Snippet: a, CLSM images of glycoRNAs by COMPASS in MCF-7 and MCF-10A cells. Scale bar, 100 μm. b, Quantification of the average fluorescence intensity of the FRET channel in a . Data in b is representative of three independent experiments; n = 5 frames. Data are shown as mean ± s.d. c, Schematic illustration of the malignancy transformation model, in which MCF-10A cells were exposed to DMBA to induce malignant transformation. d, Schematic illustration of chemotherapy model, in which MCF-7 cells were treated with PTX to induce cell death. e, Visualization of glycoRNAs abundance after treating MCF-10A cells with DMBA. Scale bar, 20 μm. f, Visualization of glycoRNAs after treating MCF-7 cells with PTX. Scale bar, 20 μm. g, Quantification of the average fluorescence intensity of the FRET channel in e . Data in g is representative of three independent experiments; n = 5 frames. Data are shown as mean ± s.d. h, The expression levels of total glycoRNA in MCF-10A cells treated with different concentrations of DMBA were assessed by MCR-free RNA blotting. i, Quantification of the average fluorescence intensity of the FRET channel in f . Data in i is representative of three independent experiments; n = 5 frames. Data are shown as mean ± s.d. j, The expression levels of total glycoRNA in MCF-7 cells treated with different concentrations of PTX were assessed by MCR-free RNA blotting. Statistical significance was analyzed by unpaired t-test; NS, not significant;(*) P < 0.05, (**) P < 0.01, (***) P < 0.001, and (****) P < 0.0001.

Article Snippet: Subsequently, DMB labeling was performed with the Sialic Acid Fluorescence Labeling Kit (Takara) and analyzed by HPLC with fluorescence detection.

Techniques: Fluorescence, Transformation Assay, Expressing

Journal: bioRxiv

Article Title: Sequence-Independent In Situ Imaging of GlycoRNA in Living Cells and Tissues based on COMPASS

doi: 10.64898/2026.04.01.715772

Figure Lengend Snippet: a, Schematic illustration of the experimental procedure for imaging mouse tissues using COMPASS. Balb/c mice (n=3) were i.v. injected with 4T1 cells in the model groups, while PBS was injected in the control group. The 5-EU (20 mg/ml) was i.p. injected initially, followed by Ac 4 ManNAz (5 mg/kg) once daily for the following three days. b, Images of the lung harvested from mice. Metastatic nodules in the model group are indicated by black circles. Scale bar, 2 mm. c, Hematoxylin & eosin (H&E) staining of tissues from the major organs of control and model mice. Metastatic nodules in the model group are indicated by black circles. Scale bar, 100 µm. d, CLSM images of FRET signals in lung tissues from different treated mice captured with the assistance of COMPASS. The experimental mice were divided into the following two groups (n = 3 per group): a control group and the model group. To establish tumor metastasis models, 4T1 cells (1×10 7 cells) suspended in PBS were injected intravenously through the tail vein in the model group. Meanwhile, the control group was injected with PBS. Mice were labeled with a single or two MCRs (100 μl of 20 mg/ml 5-EU, 0.16 mmole/kg DBCO-AF647), respectively, and subsequent slices were subjected to two-step click chemistry to achieve AF488 and AF647 labeling. Scale bar, 100 µm. e, Intensity surface plots of FRET signals in lung tissues from different treated mice captured with the assistance of COMPASS. f, Quantification of the average fluorescence intensity of nodule versus perinodule regions in the FRET channel on the right side of e . Data in f is representative of three independent experiments; n = 5 frames. Data are shown as mean ± s.d. Statistical significance was analyzed by unpaired t-test; NS, not significant;(*) P < 0.05, (**) P < 0.01, (***) P < 0.001, and (****) P < 0.0001.

Article Snippet: Subsequently, DMB labeling was performed with the Sialic Acid Fluorescence Labeling Kit (Takara) and analyzed by HPLC with fluorescence detection.

Techniques: Imaging, Injection, Control, Staining, Labeling, Fluorescence